Assessing the multimeric states of proteins: studies using laser desorption mass spectrometry.
We have developed a technique which utilizes matrix-assisted laser desorption mass spectrometry to study the subunit association of proteins. Aqueous protein samples are treated with a dilute solution of glutaraldehyde, a cross-linking agent which reacts with free amino groups on proteins. This agent effectively traps the multimeric form, preventing it from dissociating in the sample preparation and desorption process. Proteins measured include lysozyme, carbonic anhydrase, apomyoglobin, glucose 6-phosphate dehydrogenase, ovine lutropin, yeast alcohol dehydrogenase, avidin and pyruvate kinase. Dimeric and tetrameric complexes up to 250,000 Da have been measured in this manner.
Farmer
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TB
Analytical Chemistry Center, University of Texas Medical School, Houston 77030.
Caprioli
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RM
D000426:Alcohol Dehydrogenase / Q000737:chemistry; D000818:Animals; D001360:Avidin / Q000737:chemistry; D002256:Carbonic Anhydrases / Q000737:chemistry; D005954:Glucosephosphate Dehydrogenase / Q000737:chemistry; D007834:Lasers; D007986:Luteinizing Hormone / Q000737:chemistry; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods; D009113:Muramidase / Q000737:chemistry; D011506:Proteins / Q000737:chemistry*; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D012756:Sheep
D011506:Proteins; D001360:Avidin; D007986:Luteinizing Hormone; D000426:Alcohol Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; D009113:Muramidase; D002256:Carbonic Anhydrases