Protein Identification by Mass Spectrometry Issues to be Considered

摘要: During the past two decades, mass spectrometry has become established as primary method for protein identification from complex mixtures of biological origin. This is largely attributable to fortunate coincidence instrumental advances that allow routine analysis minute amounts (typically femtomoles) involatile, polar compounds such peptides in mixtures, with rapid growth genomic databases are amenable searching MS data. Like many other developing fields science, creation techniques and software tools initial generation interpretation data have been domain experts, people who cognizant not only benefits methods but also their actual potential weaknesses. Now spectrometric proteomic increasingly available accessible, a much broader range researchers applying same methodology, often substantially less understanding major limitations critically affect reliability significance results. Ideally community should establish criteria proteins be employed by researchers. As this remains rapidly field different experimental approaches ways interpreting data, it difficult promulgate hard fast rules. Nevertheless Molecular Cellular Proteomics attempting develop standards acceptability proteomics papers, based on emerging knowledge well principles over last 20 years community. Authors papers employing must make themselves fully aware key issues driving development these guidelines. Hence paper follows attempts highlight strengths weaknesses current use. It particularly important realize any match returned database search, there non-zero probability will wrong. Many times quality false positive can disregarded, some cases identifications search engines very likely incorrect. Therefore unacceptable simply list all hits come back engine then discuss though they were categorically correct. Peptide Analysis - Almost without exception, generated proteolyic digestion. The most widely used enzyme trypsin, which hydrolyses C-terminal side lysine arginine, unless subsequent amino acid sequence proline. advantageous every peptide than C-terminus at least sites efficient protonation, N-terminal group