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Re ‐face stereospecificity of NADP dependent methylenetetrahydromethanopterin dehydrogenase from Methylobacterium extorquens AM1 as determined by NMR spectroscopy
Author(s) -
Hagemeier Christoph H.,
Bartoschek Stefan,
Griesinger Christian,
Thauer Rudolf K.,
Vorholt Julia A.
Publication year - 2001
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(01)02306-7
Subject(s) - stereospecificity , chemistry , methylobacterium , nuclear magnetic resonance spectroscopy , spectroscopy , stereochemistry , methanol dehydrogenase , dehydrogenase , biochemistry , enzyme , physics , catalysis , gene , 16s ribosomal rna , quantum mechanics
MtdA catalyzes the dehydrogenation of N 5 , N 10 ‐methylenetetrahydromethanopterin (methylene‐H 4 MPT) with NADP + as electron acceptor. In the reaction two prochiral centers are involved, C14a of methylene‐H 4 MPT and C4 of NADP + , between which a hydride is transferred. The two diastereotopic protons at C14a of methylene‐H 4 MPT and at C4 of NADPH can be seen separately in 1 H‐NMR spectra. This fact was used to determine the stereospecificity of the enzyme. With (14a R )‐[14a‐ 2 H 1 ]‐[14a‐ 13 C]methylene‐H 4 MPT as the substrate, it was found that the pro‐R hydrogen of methylene‐H 4 MPT is transferred by MtdA into the pro‐R position of NADPH.