|
Carol Otey stood in front of the laboratory's cell bank freezer and pulled on a pair of asbestos gloves. Except for its institutional beige paint, the freezer looked remarkably like the Star Wars robot, R2D2. When Otey flipped the lid open a mushroom cloud of white vapor puffed into the room. As it dissipated, she used two metal rods to hoist the wire mesh basket out of the liquid nitrogen coolant.
The basket had drawers, each one containing hundreds of tiny vials full of cells being held in suspended animation. The one Otey wanted was labeled C-10 hybridoma, which was a man-made cell line embodying a cross between a mouse immune system cell and a cancer cell. These particular cells, when revived and grown out in a petri dish, should make an antibody to a specific molecule - in this case, a protein called alpha actinin.
Antbodies were the guided missiles of the immune world, evolved over the aeons to home in on foreign proteins, especially those on the surfaces of invading viruses and bacteria. Modern biology had co-oped nature by crossing cancer cells with immune system cells, creating hybrids that churned out vast quantities of antibodies to whatever protein the scientist happened to be interested in. The antibodies, once they found their targets in the cell, could be made to fluoresce under the microscope - revealing, as they did, where the target protein was and, by inference, what it was doing. In that way, the submicroscopic anatomy of life was being mapped.
Cell lines making all sorts of protein were available from biological supply houses - at a price, of course. In fact, Otey had been about to place an order for cells that produced the antibodies she needed when Keith Burridge, the head of the lab where Otey was serving as a post-doctoral student, had told her to save the money.
Some time back, the senior scientist recalled, some graduate student had made precisely the line of cells she wanted. Or he thought so, anyway, though he couldn't quite remember the project or the graduate student. Otey should look in the log book. He added, as an afterthought, that since he couldn't remember the student, neither could he vouch for the cells. So she should check them out thoroughly - make sure they were making the antibodies she wanted.
So it was that Otey came to be rummaging through the cell bank, looking for the cells the unknown student had deposited there who knows how many years ago.
The plastic vials were about an inch and a half long and were sealed with a screw cap. She found the one she wanted, retrieved it with a pair of forceps, lowered the cage back into the bubbling liquid nitrogen, and closed the top of the repository. Then she proceeded on about the business of reviving the cells, culturing them, and separating out the antibodies they made.
The project Otey was working on involved the molecular anatomy of cells. At the moment she was focusing on a protein called alpha actinin, which snapped together end-to-end to form stretchy cables that were the cellular version of muscle. She knew exactly where those proteins would be; the only question was whether the revived cells produced an antibody that would allow her to stain them.
A few days after reviving the cells, she had enough of them to extract the antibodies and use them to stain a sample of mouse cells. Then she mounted the cells on a slide, put the slide under the barrel of her microscope and adjusted the focus. The cells swam into view and, sure enough, the stain appeared precisely where the alpha actinin was supposed to be.
She would make one confirming check, which was to determine the molecular weight of whatever it was the stain was marking. That would tell the tale, because molecular weights, which were measured in daltons, roughly translated into the number of atoms in the molecules. Since it was the atomic composition that made a molecule what it was, molecular weights were definitive.
Molecules were too small to weigh individually, of course, but over the course of a century and more sophisticated solutions to that problem had evolved and been made simple and routine. By Otey's time, the process culminated in the appearance of a dark line of a piece of white paper that had been marked out with molecular weight gradiations. Otey knew from experience that alpha actinin should make a line just above the 97 kilodalton marker.
So when it appeared, instead, just short of that marker, Otey could only stare at it in disbelief.
She ran the test again, and got the same result.
It was a moment she would not forget. Night had fallen, and except for a few apprentice scientists like herself, the laboratory was deserted. She stood there, looking at the offending paper, and wrestled with the implications.
Something was wrong.
Clearly, the cells she had taken out of the nitrogen were making an antibody to some other, unknown protein. The graduate student had screwed up. But the cells were healthy enough and they were making antibodies, and the antibodies were homing in on . . . well, on something, on some protein. A protein that was not alpha actinin but was associated with it somehow, something that occurred at almost exactly the same place in the cell that the muscle-like actinin did.
The year was 1991; the place, UNC.
Otey stared at the blot, her mind forming the question that would haunt her for a decade.
What was it?
One of the peculiarities of the moment was that Otey had never wanted to be involved in such issues in the first place. Molecular biology might be the frontier of science but it just wasn't something that turned her on, thank you. In fact, growing up in the small town of Perryton in the Texas panhandle, she hadn't been able to gin up much interest in science. From what she'd seen of it, science was mainly the memorization of abstractions. The nearest she came to biology was that, as a child of the great plains, she did have a passing interest in critters.
She was a very good student, though, so when a teacher offered her extra credit if she would enter a project in the science fair, she wanted to do something with animals. She settled on an experiment to see if nutrition could have any effect on the ability of white rats to maintain their balance.
The experiment was simple. She brought home a half dozen white rats, and her father helped her make a motor-driven, rotating dowel that the animals would be enticed to walk across. She fed some of them junk food and some of them a nutritious diet and, yes, just as she expected, the well-fed rats were less likely to fall off the dowel.
The experiment won at the local science fair and took second place at the regionals in Amarillo. But the lesson she drew from it all had nothing whatsoever to do with nutrition. For while she was playing scientist her animal subjects were playing the real game of life and in the process multiplying like . . . well, like rats. By the time Otey came home from Amarillo there were several dozen of them. Her parents, good Texans that they were, drove across the state line into Oklahoma to turn the animals loose.
Today, in her laboratory at UNC, Otey remembers the experience with a rueful grin. "I guess I discovered something important. In biology, there is always something you didn't account for. Life is fundamentally unpredictable."
If memorization didn't excite her, the unpredictable did. One thing led to another and by the time she was a sophomore at Trinity University in San Antonio in the late 1970s, she was hooked on biology . . . but she retained some strong reservations. Molecular genetics was hot, and she couldn't deny its importance, but it held no thrill for her. Nor did the careful titrations of immunochemistry, or the flat, otherworldly images of electron microscopy.
No, if she was going to spend her life working out puzzles, she'd rather work on puzzles involving organisms, critters, things that lived lives of their own and did individual, interesting things. Mollusks, for example. How did those soft little animals manage to make such beautiful and infinitely varied shells? Marine biology had a definite appeal.
All the same, to be a biologist of any flavor she needed to know how the proteins and other life molecules organized themselves into cells, and how cells fitted together into living creatures. So biochemistry was required and, in the process, she became interested almost despite herself.
The molecules of life, which was to say mostly proteins, were huge things on their own scale, often consisting of hundreds of thousands of atoms and possessed of immense complexity. They had articulated parts they could turn, move, grab other things with - they became little machines that by the complexity of their function seemed capable of willful movement, in a robotic sort of way. Some of them made other molecules, some of them released energy, some of them read DNA, some of them pulled used molecules apart and sent the pieces back to the factories for recycling. The cell was a churn of chugging activity.
It was all highly organized, obviously -- there were a billion or more protein molecules in a cell, and life depended not only on each one doing the right thing, but doing it in the right place. How all this got sorted out was mysterious, but what little was known was elegant.
The biggest piece of news when Otey was in college was that each one of those proteins was turning out to come with a short address tag, coded into its surface; those tags apparently made sure everything ended up where it was supposed to be, whether in the nucleus, the cell membrane, or in one of the cell's specialized organelles. With such tags, the proteins did the work of the cell like so many well-organized craftspeople in a factory. Without them, they would have been a milling mob, and cellular life would not be possible. Before Otey had proceeded much further in her studies, in fact, researchers would reveal that several genetic diseases were caused by mis-addressed molecules.
Another hot topic in the 1970s was the discovery that cells weren't just bags of protoplasm: They had skeletons!
Like most science students Otey was fascinated. She attended a lecture on the subject and learned that the skeletons seemed to be made of two materials, one of them rigid and one of them flexible and elastic. The rigid ones, called were made of stacks of a protein; they were, in effect, the bones of the cell. The muscle analogs were made of chains of actinin molecules. Some thought the tubes elastic actinin chains worked together to form three dimensional shapes, like modern sculptures of pipes and cables; others thought it might be complicated. In truth, little was know.
Such were the facts that ended up in Otey's notebook, but what she would remember for half a lifetime was the <begin ital>experience<end ital> of science - the pervasive smells of the laboratory, the squeak of a mouse, the glint of stainless steel - the quiet thrill of unworldly precision, of manipulating things the eye could not see.
Trinity had an electron microscope that undergraduates were allowed to use, for example. Otey and her fellow students quickly discovered how much work went into using it -- and how artificial, in some respects, the result could be. When Otey's turn came with the equipment she spent days making her slide - she encased her cell sample in paraffin, shaved it thin, dried it into a chip, and fixed it on a slide.
"By the time I was done, they were no longer living things. They were dead. They were good and dead."
Even so, it was an almost religious experience to take control of the big, beige electron microscope.
The instrument was housed in a small room illuminated mainly by the otherworldly flicker of the green screen. A compressor murmured softly, and the smell of light oil hung in the air. Peering down at the screen, Otey moved the dials to control the movement of the slide. From all appearances she could have been flying a spacecraft over the surface of an alien planet . . . and yet, as her eyes adapted and she mastered the trick of piloting the hulking machine, <begin ital>there were the cells!<end ital> And they looked just like they had in the textbook, complete with membranes, organs, nuclei and all the rest.
It was shocking, almost - and it was shocking how shocking it was, given that she had known, intellectually what to expect. Biology seemed to have an endless ability to surprise and startle her. She loved that. More, she had come to doubt that she could be happy without it. She passed through the phase of wanting to be a biologist. Now she HAD to be one.
In 1980, when Otey left Texas for graduate school at UCLA, biology was entering a period of unprecedented ferment. The news in the journals was about new understanding of how genes were translated into RNA, and how RNA made its way out into the cell to serve as templates for the manufacture of proteins. Recombinant DNA was beginning.
Otey had chosen UCLA for its marine biology program, but she was beginning to have second thoughts. She wanted a life in science, and she knew that to get what you wanted, you had to be practical. To be practical, science costs money. The money wasn't there, for marine biology. But molecular biology, because of its promise to human health, was booming. It might not be her favorite thing, remote and abstract as it was, but molecular biologists had incredible toys . . .
In truth, it was all more interesting than Otey had expected. A general theory of life was beginning to grow, and with it there was emerging a hazy and theoretical - but rational and real - conception of how proteins got made and folded into their shapes, and how the result functioned. Scientists had managed to turn a few genes on and off, and while the genome project was off in the future things were moving in that direction. Somehow, though nobody really knew how, all this must inevitably translate into new understandings of how the cell worked. That, in turn, should result in treatments and cures for human diseases.
It was the dawn of a new era, and all things seem possible.
In Otey's mind a compromise grew: cell biology. Cells were important enough to draw funding, but they weren't invisible abstractions. In a certain sort of way, if you had a good imagination . . . well, you might think of them as critters.Dev 1: (continuation of previous quote) And it looked to me as if now was the time.
At UCLA, as she studied for her master's degree and then her Ph.D., news of scientific advances continued to mix with experiences that carried an emotional jolt.
There was a remarkable discovery, for example, that the tubules that made up the bones of the cell were also highways. Little motor molecules pulled themselves along them -- tractors, in practice, that grabbed a load of molecules in one part of the cell and towed it along the tubule to its destination. The first motor molecules to be discovered only went one way, but another tractor molecule was soon found that trucked along in the opposite directions. "They were like little engines on a railroad track," Otey remembers. "It was so cool." (Note to Nancy: We need one of Grey's cartoon graphics here - the same approach as for To Make a Mouse but this will be far, far simpler. I can show him just how to do it.)
It was also about that time that scientists discovered a way to visualize highly magnified cells without killing them.
"Before," Otey remembers, "if you wanted to look at what was going on inside the cell, you had to use some kind of microscopy that required that you fix the cell first. But while I was in graduate school there was this huge advance that was made in the quality of video imaging. So now it became possible to do very high resolution microscopy on live cells. You could watch them moving and you could watch structures inside them moving as well -- all this with a good degree of image quality. That was called digital enhanced video microscopy. It was developed by a guy named Bob Allen, who died a number of years ago but his wife, Nina Allen, is at NC State now. She helped with the discovery of that."
Otey's lab at UCLA didn't have the equipment to make such images at the time, however, so Otey could only look at the digital recordings made by others and envy them.
But all that was on a high intellectual plane. The life of the laboratory, which was where she spent her days, was full of experience. Even on the cellular level, she was finding out, biology was . . . life.
Early graduate school, for example, she became involved in a study of how hormones effect cells, particularly uterine cells. The research required her to scrape cells out of the uterus of a mouse and to multiply them in culture so that various hormones could be applied to them and their response observed.
Growing cells turned out to be trickier than she had expected. She had to be constantly feeding them, maintaining them, doing things for them. The smallest error, and they died. You couldn't take off for a week, when you were growing cells. It was a lot like, well . . . like gardening. Or having a pet.
Once, she had been culturing muscle cells in petri dishes, she thought she caught a movement out of the corner of her eye. She looked more closely - and what she saw would remain etched in her mind decades later.
"The cells were twitching!" she remembers in a hushed, almost reverent voice. "They were muscle cells, and they wanted to do what muscle cells do. So they started twitching."
It was not exactly a revelation, of course. She knew, intellectually, that cells were individual creatures, entities simpler but no less alive than rats or primates. But it was her cell cultures, and most particularly those muscle cells, that taught her the gut level truth of that. Once again, biology had managed to surprise her.
It was about this time that a vibrant young woman scientist, Chloe Bulinski, CQ arrived in the department. She was doing cutting edge research on the internal structures that gave cells their shape -- cell skeletons, they were called. The woman's enthusiasms and insights focused Otey's mind much as the twitching muscle cells had. With Bulinski's presence on the faculty, Otey found herself thinking more and more about the importance of cell skeletons.
Before long, Otey wanted to work in Eulinski's lab enough badly enough that she was willing to agree to do precisely the kind of work she had originally set out to avoid -- molecular chemistry. The project Bulinski assigned her focused on the nature and behavior of actinin, which made up the muscle-like fibers.
Otey was philosophical about it. "It wasn't what I had in mind, exactly, but it was good because it gave me a very strong grounding in the basic fundamental biochemical properties of the proteins that make up the cytoskeleton."
As she worked to better understand the actinin "muscles" of the cell, others around the world focused on the molecules that made up the bones - and on the large assortment of molecules that turned out to tie these and other components together inside the cell. Actinin, for example, turned out also to tie together bundles of tubules, to make them stiffer. They were applied to other functions as well - wherever the cell needed a spring or a bungee cord, there seemed to be some kind of actinin.
Otey got her Ph.D. in 1987 and moved to UNC as a post-doctoral fellow. As soon as she could, she began to move herself away from the pure biochemistry she'd had to do in California to be part of Eulinski's team. Wherever she could, now, she tried to think in terms of the whole cell.
In this way her life and her science, the two nearly indistinguishable, continued apace. Discovery led to discovery about how the various molecules made up the linkages between the cellular muscles, skeleton and other structural elements of the cell.With each passing year Otey felt more and more a worthy part of the larger scientific enterprise.
Not all was roses, though. The life of a scientist is full of failures - and those failures are often all the more frustrating because the scientist may never know the cause of the failure.
So it was that, in the routine course of her work, she needed to order a batch of antibodies to alpha actinin, and her boss told her to save the expense - look in the cryogenic freezer for the cells some other student had whipped up, long before.
From those cells she grew a colony, from the colony she harvested the antibodies and with the antibodies she labeled the mysterious protein that would be the single most frustrating discovery of her life.
Whatever the protein was, it appeared almost everywhere the actinin did. But it wasn't actinin, because actinin weighed just a hair over 97 kilodaltons, and whatever she was labeling fell just below 97.
And all she could do was stare at the result, transfixed.
The protein, whatever it was, was large as molecules go, but it was unbelievably tiny on the scale at which Otey lived her life. It was too small to see through the best of microscopes, too small to touch, too small to examine in her hands.
It was large enough, though, to lodge in her mind.
She could only stare in frustration at the lab result.
What was it?<end ital>Part 2
Carol Otey frustration over her unidentified molecule occurred against a backdrop of sober knowledge: A bench scientist's life was full of inexplicable moments, odd phenomena and unexpected results. There were lots of mystery molecule, and the odds were slim that Otey would be able to figure out what hers was.
Meanwhile, as a post-doctoral cell biologist at the University of North Carolina in the early 1990s she labored in the midst of a revolution. The molecular nature of the cell was rapidly becoming understood and its bones and muscles especially so. In those years, almost every paper on the cell skeleton seemed to drive home its potential importance not just to basic science but to human medicine as well.
The skeleton, for example, was apparently the engine of cell movement - but beyond that, the bone-and-muscle metaphor broke down. Animals moved by muscle-driven articulation of the bones, but the cell simply grew its skeleton in the direction it wanted to go. The stacks of molecules that formed the tensile structures, for example, would would dissolve on the trailing end of the cell and extend on the leading edge, pushing out the membrane and occupying new space. The springy actinin chains also dissolved where they weren't needed and grew where they were. The cell membrane flowed toward the front of the cell. Thus cells moved themselves from place to place.
That ability was fundamental to life, from the very beginning. Embryonic stem cells had to be able to navigate through the fetal tissue to reach the proper places to begin building the heart, brain, and other specialized tissues. Cells of the immune system had to seek out and attach to injured tissue. Skin cells had to be able to crawl out over scrapes and cuts.
But motility was a double-edged sword, for that was what also allowed cancer cells to metastasize from the primary tumor and spread throughout the body -- the singular quality of cancer that made it so very deadly.
It was also becoming clear that the skeleton was important not only in movement, but in adhesion as well. There was apparently a family of molecules that somehow formed a chain that attached to the skeletal tips and extended through the cell membrane. Specialized grappling molecules attached to the ends of the chain.
Those grapplers, extending through the membrane and into the outside world, were in effect the hands, feet and claws of the cell. They were the molecules, for example, that allowed wound-healing and infection-fighting cells circulating in the bloodstream to anchor themselves at the site of a wound.
As it became apparent that there were many different molecules involved in these processes - not just the handful that had been known before - there was mounting excitement in laboratories such as the one in which Otey worked. Everyone in the laboratory was working furiously to identifying new skeletal proteins and figure out the nature and function of those that were already known. The excitement made for long days, and the longest days of all were, almost by tradition, worked by graduate students and post-docs like Otey.
She had a mystery molecule of her own, of course, and she occasionally got a chance to do an experiment or two with it, but nothing worked. She knew enough to be tantalized, but not enough to justify spending time on it.
Nothing, though, could stop her from thinking about it - including Otey herself. She couldn't get it out of her mind. It intruded unbidden into her thoughts, woke her up at night.
That antibody was certainly labeling something<end ital>! But what could it be?
But if science is mysteries, then a life of science was of necessity full of questions that hung in the mind, yearning to be addressed. Otey struggled with this issue, achieving only partial success.
Aside from that frustration, though, her life was good. She liked the work, the intellectual atmosphere, the excitement of discovery. And she liked the aesthetics of it, as well.
No matter how many times she saw the moving digital images of cells crawling across cover slips, she never grew bored. Her life was full of such images, skeletons fluorescing, molecules streaming, skeletal members being stripped away on one side and built up on the other, cell membranes streaming. It was all very mechanistic, she knew in her mind; structures were built up here, broken down there . . . it could all be reduced to molecular explanation. Yet all that construction and deconstruction happened very rapidly, as if time, on that small scale, had been dramatically accelerated, and if the cell was made of molecular robots, it certainly was not one itself. It moved gracefully, with a fluid motion, its forward parts ruffling delicately as it felt its way, exploring virgin territory.
The fledgling scientist that was Carol Otey worked away, rigging experiments, tending petri dishes, making technical observations about the moving cell and filing them away. She was good at it. Precise. Intellectual. Reductionistic.
But the human being in her could not help but notice the other thing, as well.
Cells were creatures, all right. But not only that. They were also . . . beautiful.
The most cherished dream of most young scientists is to secure a faculty position at a teaching university, but when Otey was hired to set up a laboratory at the University of Virginia in 1993, the achievement was bittersweet because - not long before that, the perfect man had walked into her life.
His name was Aldo Rustioni. He was an Italian scientist who she knew vaguely from meetings. She thought he had a cute accent. Then one day he wandered into her lab, shuffling through a handful of papers. He apologized, claiming that in his preoccupation he had gotten off the elevator on the wrong floor, but before he left the lab Otey had agreed to join him for dinner. Before she left for the University of Virginia, they were married, and Otey found herself involved in a commuter marriage.
It took her almost four hours to make the trip, and as the next few years passed they both got to know the route well. Both looked for a job at the other's university, but faculty science positions are not simple to secure. So they used part of each weekend to schmooze at whatever campus they were near.
The other difficulty in her life was that there was no progress whatsoever on the mystery protein. When she left UNC, Burridge told her she might as well take antibody-producing cells with her; technically speaking, they were his, but she was far more likely to do something with them than he was.
But though the cells were never far from her mind, the life of a scientist was driven by the need to do something that applied right now<end ital>, that would get a paper in the journals, that would apply to the next grant application. So while she kept promising herself she'd spend some time on the mystery, her days now seemed even fuller than they did at UNC. So the problem was strictly back burner . . . no, it wasn't even that. Instead of sitting in Burridge's liquid nitrogen, they set in hers.
Other than that, though, Otey's science was booming. At UVA she quickly began to collect graduate students, among them one Magnus Edlund. Edlund, like Otey was fascinated by the idea that super-cooled microscopes linked with computers could take digital moving pictures of live cells. He wanted to build such an instrument as his project and Otey was quick to okay his proposal - perhaps not entirely for scientific reasons. Edlund spent the next months of his life working in a tiny, dim room, his face lit only by the sickly green of cathode ray tubes, tinkering with computer chips and liquid nitrogen baths.
Then one day in 1996 Edlund appeared at Otey's side, flushed with excitement. She ran down the hallway on his heels and burst into the little room. She bent over the microscope, pressed her face against the eyepieces and . . .
There it was, a living cell, making its way across the field of view, feeling its way forward with delicate ruffles of protoplasm, grasping the surface of the cover slip with invisible claws . . . it was the experience of a lifetime.
"We were there for hours," she would remember later. "First I'd look, then he'd push me away so he could look. Then I'd push him away so I could look. So we took turns like that.
So as the years passed, Otey settled into the hectic life of the modern scientist, supervising graduate students, writing papers and grants, attending meetings and taking part in the insider's intellectual gossip that is the cultural requirement of the scientific endeavor. The mystery cells sat patiently in the liquid nitrogen.
Then one day she was sitting in her office with a new MD-Ph.D. student, Mana Parast., discussing research possibilities her. Parast was a tiny woman who sat on the edge of her chair and seemed intensely interested in every possibility Otey offered to her.
Then, subliminally encouraged by Parast's rapt attention, Otey found herself telling the story of the mystery cells, and how their antibodies had latched onto what was apparently an unknown protein.
Halfway through the exposition something in Parast's manner, or the rapt expression on her face, prompted Otey to have second thoughts. It was important that a graduate student undertake something that would succeed. Their first research project should reward them with a paper, not tie them up in knots. And that protein, whatever it might be, had the earmarks of a bottomless intellectual pit. Perhaps another project . . .
But Parast wouldn't hear it. What she wanted to know was where the cells were, and when she could get started.
Otey's studied the woman sitting across from her. She, Otey, had worked in science long enough to recognize an irresistible force when she saw it. Knowing she had no choice, she acquiesced.
Then one Friday her husband called her in Virginia and said he wasn't feeling well. It was his turn to commute but . . . would she mind driving south instead? She agreed and then, before she left, she called him back - and hardly recognized his voice.
"He was having shortness of breath, tightness of the chest, all the classic symptoms. He was terrified. He had called the paramedics, and they were on way.
She made the trip to UNC in three hours flat. She went directly to the hospital, where he was in the coronary care unit and the cardiology team was scheduling him to have stents [cq] put in two of his arteries.
In the days that followed, Otey learned all about stents, which were essentially hollow tubes put in place in an artery to hold it open. It worked . . . but there was one major problem. In a significant number of cases, cells from the arteries crawled into the stent and ultimately filling the life-giving channel with cells.
As Otey waited to find out if that would happen in her husband's case, she acquired a whole different view of cell motility. Her piece of the biological puzzle was important to real people in the real world -- and, as always, biology was unpredictable. In this case, far too unpredictable.
In the event, her husband's stents remained clear. But she saw her job in a different light - and she also redoubled her efforts to find a job closer to him. Finally, in 1998, she moved her laboratory to UNC.
The thrust of her work, at the time, involved prostate cancer. Many such cancers remained in place, and didn't metastasize, for years. But others produced motile cells early and, as a result, often spread before the tumor could be diagnosed. Otey was trying to find out what triggered that early motility.
But she also kept in close touch with Parast, who remained at UVA and was working with the mystery cells.
In modern science, with fax machines, internets and dirt-cheap long distance calls, geography is not as important that it used to be. Though Otey was now separated from her graduate student by a three-hour stretch of highway, they continued their collaboration on an almost daily basis. About once a month Parast would drive down to UNC to work a day or two in Otey's lab.
Slowly, methodically, Otey and Parast used the tricks and instruments of modern molecular biology to narrow the possibilities as to what the mystery protein might be. Each experiment seem to confirm that, whatever it was, it was involved in the cell's architecture. Ultimately, convinced it was a previously unknown protein, they named it Palladin for the Renaissance architect Andreo Pallardio, whose work spread across America in the 1700s through the fertile medium of Thomas Jefferson.
Then, in October of 1999, something happened electrified cell biologists around the globe. Guenter Blobel, the man who discovered that all the molecules in the cell had their own address tag, won the Nobel Prize.
"We were jubilant," Otey says."This was the first time a cell biologist had won a Nobel. We talked about almost nothing else for days. It was like the whole field, all of cell biology, had won something. And I think it occurred to me, then, for the first time, that I had somehow . . . that cell biology used to be an obscure field, relatively speaking. But it no longer is. Now it's the cutting edge."
For thirty years molecular genetics had reigned supreme in biology generally and even in science as a whole, the way physics had dominated the early part of the century. Now it was bearing fruit as genome after genome was sequenced: yeast, e. coli, nematodes (roundworms) and fruit flies. The human genome project was nearing fruition, with the genome of the laboratory mouse not far behind.
But if the code of life was being translated, the meaning of what it said was far from clear. How did the code specify for the processes of life - and what, for that matter, were those processes? What were the proteins, and how did they work so that life would happen, that glandular cells would secrete hormones, gut cells would secrete gastric juices, that kidney cells would purify the blood and brain cells would charge and discharge their impulses . . . and that immune system cells would move, and heart cells would stay put? That, now, in microcosm, was the question Otey and Parast were asking.
But was their question apt, and were they asking it correctly?
By October of 1999 Otey and Parast had a partial sequence for the mystery protein, which revealed it was indeed an unknown one. That, finally, made it possible to do the critical experiment: They would disable Palladin in a cell culture, and see how the creatures performed without it.
This would be the moment of truth. Cells were full of proteins, and the lore of science was full of luckless researchers who spent great amounts of time, effort and money to isolate a protein only to find that, when they knocked it out . . . nothing happened. There was no effect at all. The cell went on about its business. The protein they worked so hard to characterize was a byproduct of some sort, or had a function that was duplicated by many other proteins.
But then, of course, sometimes when a mystery protein is knocked out, something dramatic happens . . . and the result was not lore but history.
Soon, they would know.
The experiment was done one day in November of 1999. Parast did it, in Virginia.
Otey will never forget the moment when, working in her laboratory, she reached over to pick up the ringing phone. Parast was on the other end of the line, bubbling ecstatically about profound changes in the cells. When the Palladin proteins of a cell were neutralized, the cells lost the ability to move! They couldn't do anything! They just drew up into balls and floated there!
Otey stood, transfixed, at the telephone.
In that case Paladin . . . whatever it did, whatever its still-mysterious function, it was fundamental to life.
"I thought, wow! But then I thought whoa! I couldn't quite bring myself to believe it. In all likelihood, there had been a mistake. This was something I had to see with my own eyes."
She ordered Parast to pack what she needed and drive down to UNC -- right now, immediately. They'd repeat the experiment, together.
It took the better part of the following day to set up and run the experiment. Otey insisted on double-checking everything, controls for every possible variation, every caution she could think of. But when was over, the results were the same.
"The entire shape of the cell had been lost. It just rounded out up. This was just amazing.
"Mona and I . . . I mean, we were jumping up and down. Literally jumping up and down."
The implications, only partly formed, swirled through Otey's mind.
Without Palladin, the whole idea of the organization life, of the organism itself, was impossible
If the molecule was that important, some small genetic or environmentally induced change in it might change the cells in important ways; who knows how many diseases might be related to problems with palladin?
Cancer, when it metastasized, probably used the palladin molecule -- metastasis was what happened when a cell that wasn't supposed to be able to move any more suddenly regained that ability, slipped into the blood stream, and found a home someplace else in the body. Did that imply that palladin was involved in the gensis of cancer or, alternately, might the palladin in cancer cells somehow be disabled to halt metastasis?
Who could know?<end ital>
All that was but speculation, of course, wisps of possibilities that ran through the mind, fueled by equal measures of exhaustion and adrenaline. But from that distillate was formed the intoxicant that powers science.
The following weeks were intense. Otey dropped the prostate cancer work and reorganized her laboratory around Palladin. Meanwhile she and the graduate student began writing a long paper announcing the discovery and providing the description of the new molecule. She told some friends and colleagues what had happened, and the gossip began to spread. The Journal of Cell biology snapped up the scientific paper and published it in August.
"And so that's where we're at," says Otey, some of the marvel of it all caught in her voice. "Everything we learn about the genome has to be worked out, ultimately, in the cell. That's going to be the news in the next few decades . . . and, wow, here I am. Am I surprised? Yes!
"The biggest surprise of all to me is how interesting Palladin has turned out to be. I remember how, when we first started working on it, we weren't even really sure it was new . . . my husband kids me about how for years I'd come home every day and worry about how the project was going to completely fall apart and go bust . . . that, or it'd turn out to be interesting. But I never thought it'd be THIS interesting."
She pauses for a moment, reflecting on the road she traveled, from white rats to twitching cells to a strange molecule that now, somewhere in its mysterious nature, bears her future as a scientist.
"I guess that's the lesson of biology," she said. "Life is unpredictable. You just never can tell." |
|
