Tryptophan aminopeptidase activity of several indole prenyltransferases from Aspergillus fumigatus.
Recently, five indole prenyltransferases from Aspergillus fumigatus have been proven biochemically to be responsible for prenylations of diverse substrates. In this study, we show peptidase activities of 7-DMATS, FgaPT1, CdpNPT, and FtmPT1, with preference for linear peptides containing a tryptophanyl moiety at the N terminus. Testing of 31 peptides revealed that these enzymes shared similar substrate specificity and accepted H-L-Trp-L-Ala-OH and H-L-Trp-Gly-OH as best substrates for aminopeptidase activity. By using H-L-Trp-Gly-OH as substrate, Km values at 350, 380, 300, and 420 microM and enzymatic rate constants kcat/Km at 0.51, 0.24, 0.53, and 0.14 mM(-1)s(-1) were determined for 7-DMATS, FgaPT1, CdpNPT, and FtmPT1, respectively. In contrast to prenyltransferase activities, the aminopeptidase activities were strongly or completely inhibited by EDTA. Mn2+ increased the aminopeptidase activities of FtmPT1 and CdpNPT up to 4- and 6-fold, respectively. To the best of our knowledge, this is the first report on the catalytic promiscuity of prenyltransferases.
Kremer
Anika
A
Heinrich-Heine-Universität Düsseldorf, Institut für Pharmazeutische Biologie und Biotechnologie, Universitätsstrasse 1, D-40225 Düsseldorf, Germany.
Li
Shu-Ming
SM
D000595:Amino Acid Sequence; D000626:Aminopeptidases / Q000737:chemistry*; D001232:Aspergillus fumigatus / Q000201:enzymology* / Q000378:metabolism; D004122:Dimethylallyltranstransferase / Q000737:chemistry*; D004492:Edetic Acid / Q000737:chemistry; D006868:Hydrolysis; D007211:Indoles / Q000737:chemistry*; D007700:Kinetics; D008345:Manganese / Q000737:chemistry; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry; D011994:Recombinant Proteins / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D014364:Tryptophan / Q000737:chemistry*
D007211:Indoles; D010455:Peptides; D011994:Recombinant Proteins; D008345:Manganese; D014364:Tryptophan; D004492:Edetic Acid; D004122:Dimethylallyltranstransferase; D000626:Aminopeptidases