Detecting native folds in mixtures of proteins that contain disulfide bonds.
High-throughput in vitro refolding of proteins that contain disulfide bonds, for which soluble expression is particularly difficult, is severely impeded by the absence of effective methods for detecting their native forms. We demonstrate such a method, which combines mass spectrometry with mild reductions, requires no prior experimentation or knowledge of proteins' physicochemical characteristics, function or activity, and is amenable to automation. These are necessary criteria for structural genomics and proteomics applications.
Narayan
Mahesh
M
Baker Laboratory of Chemistry and Chemical Biology, Cornell University, Ithaca, New York 14353-1301, USA. mnarayan@utep.edu
Welker
Ervin
E
Zhai
Huili
H
Han
Xuemei
X
Xu
Guoqiang
G
McLafferty
Fred W
FW
Scheraga
Harold A
HA
D000465:Algorithms; D001665:Binding Sites; D045424:Complex Mixtures / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry*; D010449:Peptide Mapping / Q000379:methods*; D011485:Protein Binding; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*
D045424:Complex Mixtures; D004220:Disulfides; D011506:Proteins