Mass processing--an improved technique for protein identification with mass spectrometry data.
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis following tryptic digestion of polyacrylamide gel pieces is a common technique used to identify proteins. This approach is rapid, sensitive, and user friendly, and is becoming widely available to scientists in a variety of biological fields. Here we introduce a simple and effective strategy called "mass processing" where the list of masses generated from a mass spectrometer undergoes two stages of data reduction before identification. Mass processing improves the ability to identify in-gel tryptic-digested proteins by reducing the number of nonsample masses submitted to protein identification database search engines. Our results demonstrate that mass processing improves the statistical score and rank of putative protein identifications, especially with low-quantity samples, thus increasing the ability to confidently identify proteins with mass spectrometry data.
Henkin
Josh A
JA
Department of Biology, The University of Vermont, Burlington, USA. Joshua.Henkin@uvm.edu
Jennings
Mark E
ME
Matthews
Dwight E
DE
Vigoreaux
Jim O
JO
D002623:Chemistry Techniques, Analytical; D030562:Databases, Protein; D004591:Electrophoresis, Polyacrylamide Gel; D011506:Proteins / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D011506:Proteins