On the role of tryptophan residues in the mechanism of action of glyceraldehyde-3phosphate dehydrogenase as tested by specific modification.
A method is described to selectively modify one of the three tryptophan residues of the subunit of glyceraldehyde-3-phosphate dehydrogenase from yeast.  As modifying agent dimethyl (2-hydroxy-5-nitrobenzyl) sulfonium bromide was used.  The residue which is modified by the procedure described has been identified as Trp-193.  There are either one or two molecules of the modifying agent being added to this tryptophan side chain.  The modification apparently does not cause a detectable conformational change of the protein as judged from the methods employed.  However, the enzymatic activities in the dehydrogenase as well as in the esterase reactions are lost after the modification.  It could be established that the modification rendered the enzyme unable to bind the oxidized coenzyme.  Also the charge-transfer interaction between enzyme and coenzyme could no longer be observed.
D001059:Apoproteins; D001593:Benzyl Compounds / Q000494:pharmacology; D001665:Binding Sites; D002850:Chromatography, Gel; D002918:Chymotrypsin; D003956:Dialysis; D004589:Electrophoresis, Disc; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D007700:Kinetics; D009243:NAD; D009574:Nitro Compounds / Q000494:pharmacology; D011485:Protein Binding; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D013438:Sulfhydryl Compounds / Q000032:analysis; D013452:Sulfonium Compounds / Q000494:pharmacology; D014357:Trypsin; D014364:Tryptophan / Q000032:analysis*
D001059:Apoproteins; D001593:Benzyl Compounds; D009574:Nitro Compounds; D013438:Sulfhydryl Compounds; D013452:Sulfonium Compounds; D009243:NAD; D014364:Tryptophan; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases; D002918:Chymotrypsin; D014357:Trypsin