Catalytic mechanism of stereospecific formation of cis-configured prenylated pyrroloindoline diketopiperazines by indole prenyltransferases.
Indole prenyltransferases AnaPT, CdpC3PT, and CdpNPT are known to catalyze the formation of prenylated pyrroloindoline diketopiperazines from tryptophan-containing cyclic dipeptides in one-step reactions. In this study, we investigated the different stereoselectivities of these enzymes toward all the stereoisomers of cyclo-Trp-Ala and cyclo-Trp-Pro. The stereoselectivities of AnaPT and CdpC3PT mainly depend on the configuration of the tryptophanyl moiety in the substrates, and they usually introduce the prenyl moiety from the opposite sides. CdpNPT showed lower stereoselectivity, and the structure of the second amino acid moiety in the substrates is important for the stereospecificity in its enzyme catalysis. Moreover, we determined the crystal structure of AnaPT in complex with thiolodiphosphate and compared it with the known structures of CdpNPT. Our results clearly revealed the presence of an indole binding mode that has so far not been characterized.
Institut für Pharmazeutische Biologie und Biotechnologie, Philipps-Universität Marburg, Marburg 35037, Germany.
Xia
X
Yu
Interfakultäres Institut für Biochemie, Eberhard Karls Universität Tübingen, Tübingen 72076, Germany.
Georg
G
Zocher
Fachbereich Chemie, Philipps-Universität Marburg, Marburg 35032, Germany.
Xiulan
X
Xie
Institut für Pharmazeutische Biologie und Biotechnologie, Philipps-Universität Marburg, Marburg 35037, Germany.
Mike
M
Liebhold
Interfakultäres Institut für Biochemie, Eberhard Karls Universität Tübingen, Tübingen 72076, Germany.
Stefan
S
Schütz
Interfakultäres Institut für Biochemie, Eberhard Karls Universität Tübingen, Tübingen 72076, Germany. Electronic address: thilo.stehle@uni-tuebingen.de.
Thilo
T
Stehle
Institut für Pharmazeutische Biologie und Biotechnologie, Philipps-Universität Marburg, Marburg 35037, Germany. Electronic address: shuming.li@staff.uni-marburg.de.
Shu-Ming
SM
Li
D001232:Aspergillus fumigatus / Q000737:chemistry / Q000201:enzymology* / Q000378:metabolism; D055162:Biocatalysis; D018360:Crystallography, X-Ray; D054659:Diketopiperazines / Q000737:chemistry / Q000378:metabolism*; D004122:Dimethylallyltranstransferase / Q000737:chemistry / Q000378:metabolism*; D004151:Dipeptides / Q000737:chemistry / Q000378:metabolism; D007211:Indoles / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D055326:Neosartorya / Q000737:chemistry / Q000201:enzymology* / Q000378:metabolism; D010456:Peptides, Cyclic / Q000737:chemistry / Q000378:metabolism*; D054876:Prenylation; D011392:Proline / Q000031:analogs & derivatives / Q000737:chemistry / Q000378:metabolism; D013237:Stereoisomerism; D013379:Substrate Specificity
D054659:Diketopiperazines; D004151:Dipeptides; D007211:Indoles; D010456:Peptides, Cyclic; C113296:tryptophyl-proline; C057812:indoline; C030374:indole; D011392:Proline; D004122:Dimethylallyltranstransferase