PMID- 6104660
OWN - NLM
STAT- MEDLINE
DCOM- 19800928
LR  - 20190511
IS  - 0021-924X (Print)
IS  - 0021-924X (Linking)
VI  - 87
IP  - 5
DP  - 1980 May
TI  - Studies on the reaction of D-amino acid oxidase with beta-cyano-D-alanine. 
      Observation of an intermediary stable charge transfer complex.
PG  - 1469-81
AB  - The reaction of D-amino acid oxidase [EC 1.4.3.3] (DAO) from porcine kidney with 
      beta-cyano-D-alanine (D-BCNA) was studied. DAO was found to catalyze elimination 
      of the cyano group as well as oxidation of D-BCNA. During the course of the 
      reaction in the presence of excess oxygen, an intermediate was observed which 
      exhibited a characteristic absorption spectrum with a broad charge transfer band 
      in the longer wavelength region. The CD spectrum of this intermediate resembles 
      that of DAO-anthranilate complex. The rate of oxygen consumption in the aerobic 
      reaction decreased with time, suggesting product inhibition due to complex 
      formation between the enzyme and the product. Anaerobic addition of D-BCNA 
      reduced the enzyme to its fully reduced state, the CD spectrum of which closely 
      resembles that of the enzyme reduced by excess D-alanine. When an appropriate 
      amount of D-BCNA was added to the enzyme under air, the charge transfer complex 
      was observed immediately, and underwent a change to the reduced state as the 
      oxygen was consumed. The binding strength in the charge transfer complex was 
      found to be comparable to that in DAO-benzoate complex. The accumulating product 
      in the oxidation of D-BCNA had a strong absorption at 285 nm. The aerobic 
      reaction of beta-cyano-L-alanine (L-BCNA) with snake venom L-amino acid oxidase 
      (LAO) produced the same product with an absorption at 285 nm as the reaction of 
      DAO with D-BCNA. The product obtained in the reaction with LAO was found to form 
      the same charge transfer complex with DAO. We tentatively identified this product 
      as alpha-amino-beta-cyanoacrylate and the charge transfer complex as the complex 
      of alpha-amino-alpha-cyanoacrylate with the oxidized enzyme. A hypothetical 
      reaction pathway based on the present finding is proposed. Addition of L-BCNA to 
      the enzyme produced an absorption spectrum very similar to that of the 
      DAO-benzoate complex without oxidation or elimination. L-BCNA was found to be a 
      competitive inhibitor of the oxidation of D-alanine.
FAU - Miura, R
AU  - Miura R
FAU - Shiga, K
AU  - Shiga K
FAU - Miyake, Y
AU  - Miyake Y
FAU - Watari, H
AU  - Watari H
FAU - Yamano, T
AU  - Yamano T
LA  - eng
PT  - Journal Article
PL  - England
TA  - J Biochem
JT  - Journal of biochemistry
JID - 0376600
RN  - 0 (Cyanides)
RN  - 923-01-3 (3-cyanoalanine)
RN  - EC 1.4.3.3 (D-Amino-Acid Oxidase)
RN  - OF5P57N2ZX (Alanine)
SB  - IM
MH  - Alanine/*analogs & derivatives
MH  - Animals
MH  - Circular Dichroism
MH  - Cyanides
MH  - D-Amino-Acid Oxidase/*metabolism
MH  - Kidney/enzymology
MH  - Kinetics
MH  - Oxygen Consumption
MH  - Protein Binding
MH  - Protein Conformation
MH  - Spectrophotometry
MH  - Swine
EDAT- 1980/05/01 00:00
MHDA- 1980/05/01 00:01
CRDT- 1980/05/01 00:00
PHST- 1980/05/01 00:00 [pubmed]
PHST- 1980/05/01 00:01 [medline]
PHST- 1980/05/01 00:00 [entrez]
AID - 10.1093/oxfordjournals.jbchem.a132888 [doi]
PST - ppublish
SO  - J Biochem. 1980 May;87(5):1469-81. doi: 10.1093/oxfordjournals.jbchem.a132888.