Analysis of non-covalent protein complexes up to 290 kDa using electrospray ionization and ion trap mass spectrometry.
Non-covalently-bound subunit complexes of proteins have been measured by an ion trap mass spectrometer equipped with an orthogonal electrospray ionization source. For the analysis of the generated molecular ions with high mass/charge ratios, the mass/charge range of the ion trap was extended by increasing its radio frequency (rf) voltage to 15 kV (V(0-p)) and by resonant ion ejection. Ions of the non-covalent dimer of bovine serum albumin (BSA), as well as of subunit complexes of alcohol dehydrogenase (ADH) from bakers' yeast and from horse liver, have been detected at mass/charge values between 3000-9000 Th. The maximum observed molecular weight was that of a non-covalently-bound subunit-octamer of bakers' yeast ADH (two non-covalently-bound subunit-tetramers) at ca. 290 kDa.
Wang
Y
Y
Bruker-Daltonik GmbH, Fahrenheitstrasse 4, 28359 Bremen, Germany. yangwang@magnet.fsu.edu
Schubert
M
M
Ingendoh
A
A
Franzen
J
J
D000426:Alcohol Dehydrogenase / Q000032:analysis* / Q000737:chemistry; D000818:Animals; D002417:Cattle; D019281:Dimerization; D005656:Fungal Proteins / Q000032:analysis / Q000737:chemistry; D006736:Horses; D008099:Liver / Q000201:enzymology; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods*; D012441:Saccharomyces cerevisiae; D012710:Serum Albumin, Bovine / Q000032:analysis* / Q000737:chemistry
D005656:Fungal Proteins; D012710:Serum Albumin, Bovine; D000426:Alcohol Dehydrogenase