How do you conduct a research on the molecular structures of proteins like p53 in a lab?

I'm a senior in high school about to graduate in a few months, I plan to do a major research for my senior final involving the study of p53 and its affects on cancer growth. Ideally, I would need the help of a local university professor (SFSU) although I want to be sure what I'm doing so I could have a little more confidence with how to deal with state-of-the-art lab materials. What are the equipments used to study molecular proteins? How do you use it and finally analyze the results? Anyone with a Life Science background are more than welcomed to contribute! :-)

How do you conduct a research on the molecular structures of proteins like p53 in a lab?
p53 is probably the most studied protein with well over 40000 scientific papers about it. It is very brave of you to intend to familiarize yourself with this subject and make a contribution to the field in half a year that you have before graduating. From your question it appears that you have a very long way to go.


p53 structure is considered known for a decade or so from crystallography studies (which require equipment worth millions of $$$), but, I suppose, new aspects of it could still be discovered. The functional studies would depend on the type of question being asked. Typical tissue culture work would require at least a laminar hood, and incubator and a microscope, plus pipettes, plates, growth medium, etc. The hardware alone will be in tens of thousands. I hope you do not plan to set up your own lab.


Some important questions could be (and have been) asked on patient samples (e.g. mutations in cancer; role in response to therapy; etc.). This may be a bit cheaper to do, but the access to patient samples is very strictly regulated.


If you intend to do some real research you would be better off joining a well-established lab and learning what kind of work is going on there, than inventing a project all by yourself. Good luck!
Reply:p53 is as someone already pointed out very intensely studied and has been sequenced by proteomics experts. p53 has been known as "the guardian of the genome", due to its crucial role in regulating the cell cycle, in particular that it can halt mitosis until damaged DNA or mismatched bases can be repaired, thereby preventing infidel DNA from being passed onto a daughter cell (also, if the DNA damage can't be repaired, p53 can direct the cell to undergo apoptosis, or cell suicide, which also prevents infidel DNA from being passed to future generations).





With regard to its significance to cancer... the gene that encodes the 53kDa protein - p53, has been shown to carry mutations in critical regions in some 50 percent of cancers. A mutated p53 protein can't perform the functions of cell cycle arrest or induction of apoptosis... therefore, cells with a non-functional p53 are much more likely to escape regulated apoptosis, and should they be cancerous cells, tumour growth is a lot more likely. It should be noted however that a lot of other growth promoters and growth inhibitors have an integral role in cellular proliferation, eg zymogens such as the family of caspases, and p53 is not solely responsible for these processes.





Equipment and consumable materials associated with cell and tissue culture studies are indeed expensive, and as such, you would require a facility that would permit you to perform your studies there - having said that, without a degree in a life science and some heaughty funding or sponsorship behind you, I can't imagine any place just letting you walk in the door.





The primary techniques currently used in studying proteins all involve a technique called gel electrophoresis. You can look that up online for yourself, however, i can give you a simple example of what a typical approach might be...





A suspected cancerous growth would have its DNA extracted and amplified using polymerase chain reaction (PCR - also look this up online, simple enough). The target gene p53, is what is amplified using specific primers. A normal p53 gene serves as the control and is also amplified via PCR. A technique called PCR-SSCP (look it up!) can identify if a mutation is present in the suspect targeted gene from the possible tumour. Both of the oligonucleotides are analysed on a polyacrylamide gel electrophoretically. A single mutation in p53 in the possible tumour growth would result in it having a different electrophoretic mobility and as such, it would migrate at a different rate in the gel than the non-mutated, normal p53. That's genetic analysis... other methods involve studying the actual protein p53 rather that the gene that it is transcribed from - proteomics.