Peptide-mass profiles of polyvinylidene difluoride-bound proteins by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry in the presence of nonionic detergents.
Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), in conjunction with enzymatic digestion of proteins and molecular weight search of peptide-mass database is a powerful technique for peptide/protein identification. Ideally, peptide mixtures should be compatible with both MALDI-TOF and microsequencing. In our laboratory, enzymatic digestion and extraction of peptides from polyvinylidene difluoride (PVDF)-bound proteins is performed in the presence of nonionic detergents. However, nonionic detergents have been shown to cause signal suppression in MALDI-TOF analysis. This study demonstrates that by using a modified matrix solution, peptide-mass fingerprinting of PVDF-bound proteins by MALDI-TOF can be obtained in the presence of nonionic detergents such as hydrogenated Triton X-100 (RTX-100), octylglucopyranoside, and Tween 20.
Gharahdaghi
F
F
Rockefeller University Protein/DNA Technology Center, New York, New York 10021, USA.
Kirchner
M
M
Fernandez
J
J
Mische
S M
SM
D000595:Amino Acid Sequence; D003902:Detergents; D005960:Glucosides; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D017830:Octoxynol; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D010449:Peptide Mapping; D011136:Polysorbates; D011145:Polyvinyls; D011485:Protein Binding; D011506:Proteins / Q000032:analysis / Q000737:chemistry*; D012996:Solutions; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014168:Transferrin / Q000032:analysis / Q000737:chemistry / Q000235:genetics; D005465:Fluorocarbon Polymers
D003902:Detergents; D005960:Glucosides; D017830:Octoxynol; D010446:Peptide Fragments; D011136:Polysorbates; D011145:Polyvinyls; D011506:Proteins; D012996:Solutions; D014168:Transferrin; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers; C018619:octyl-beta-D-glucoside