MadSci Network: Molecular Biology |
Hello, These days, the amino-acid sequence of a protein is inferred by the nucleic acid sequence. Sequencing a DNA is indeed far more easier than sequencing a protein. Once you know the DNA sequence, usually there is no problem knowing the protein squence. For the technique concerning DNA sequencing, I can advise you to read here : http://www.bio.davidson.edu/Courses/Molbio/MolStudents/spring2003/Obenrader/sanger_method_page.htm The genetic code is univocal knowing the organism you work with : http://molbio.info.nih.gov/molbio/gcode.html To answer your question, there are indeed ways to know directly the sequence of a protein. First the protein is analysed interms of weight and composition. This is done by running the protein in acrylamide gels : http://en.wikipedia.org/wiki/SDS-PAGE Then there are two techniques : 1) the chemical (and old) one, which is, to my knowledge, never used anymore these times : it is called the Edman reaction : with a certain chemical, you digest (remove) the last amino-acid of a chain, which is then identified (liquid chromatography). Then you repurify your protein and you do the reaction again, and this provides you the nature af the second amino-acid. This is pure organic chemistry : http://en.wikipedia.org/wiki/Edman_degradation 2) Mass spectrometry : here you have two possibilities (in terms of ionisation) : MALDI and electrospray. The protein is first digested with different endoproteases (trypsin for exemple), then the peptides are ionised, selected and fragmented. The filiation of the fragments provides you the sequence. For the technical explanation (which are very difficult) about mass spectrometry I advise you this site : http://en.wikipedia.org/wiki/Mass_spectrometry#Mass_spectrometry_of_proteins Hope this helps See you...
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