| Title | Identification of 2D-gel proteins: a comparison of MALDI/TOF peptide mass mapping to mu LC-ESI tandem mass spectrometry. |
| Publication Type | Journal Article |
| Year of Publication | 2003 |
| Authors | Lim H, Eng J, Yates JR, Tollaksen SL, Giometti CS, Holden JF, Adams MWW, Reich CI, Olsen GJ, Hays LG |
| Journal | J Am Soc Mass Spectrom |
| Volume | 14 |
| Issue | 9 |
| Pagination | 957-70 |
| Date Published | 2003 Sep |
| ISSN | 1044-0305 |
| Keywords | Amino Acid Sequence, Chromatography, Liquid, Databases, Protein, Electrophoresis, Gel, Two-Dimensional, Methanococcus, Molecular Sequence Data, Molecular Weight, Peptide Mapping, Proteins, Pyrococcus furiosus, Sensitivity and Specificity, Software, Spectrometry, Mass, Electrospray Ionization, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Trypsin |
| Abstract | A comparative analysis of protein identification for a total of 162 protein spots separated by two-dimensional gel electrophoresis from two fully sequenced archaea, Methanococcus jannaschii and Pyrococcus furiosus, using MALDI-TOF peptide mass mapping (PMM) and mu LC-MS/MS is presented. 100% of the gel spots analyzed were successfully matched to the predicted proteins in the two corresponding open reading frame databases by mu LC-MS/MS while 97% of them were identified by MALDI-TOF PMM. The high success rate from the PMM resulted from sample desalting/concentrating with ZipTip(C18) and optimization of several PMM search parameters including a 25 ppm average mass tolerance and the application of two different protein molecular weight search windows. By using this strategy, low-molecular weight ( |
| Alternate Journal | J. Am. Soc. Mass Spectrom. |
| PubMed ID | 12954164 |
| Grant List | RR-11823 / RR / NCRR NIH HHS / United States |
Department of Microbiology