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Mechanism of interaction of O-amino-D-serine with sheep liver serine hydroxymethyltransferase.

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Title Mechanism of interaction of O-amino-D-serine with sheep liver serine hydroxymethyltransferase.
 
Creator Baskaran, N.
Prakash, V.
Appu Rao, A. G.
Radhakrishnan, A. N.
Savithri, H. S.
Appaji Rao, N.
 
Subject 07 Enzyme Biochemistry
Meat
 
Description The mechanism of interaction of O-amino-D-serine (OADS) with sheep liver serine hydroxymethyltransferase (EC 2.1.2.1) (SHMT) was established by measuring changes in the enzyme activity, absorption spectra, circular dichroism (CD) spectra, and stopped-flow spectrophotometry. OADS was a reversible noncompetitive inhibitor (Ki = 1.8 microM) when serine was the varied substrate. The first step in the interaction of OADS with the enzyme was the disruption of enzyme-Schiff base, characterized by the rapid disappearance of absorbance at 425 nm (6.5 X 10(3) M-1 s-1) and CD intensity at 430 nm. Concomitantly, there was a rapid increase in absorbance and CD intensity at 390 nm. The spectral properties of this intermediate enabled its identification as pyridoxal 5'-phosphate (PLP). These changes were followed by a slow unimolecular step (2 X 10(-3) s-1) leading to the formation of PLP-OADS oxime, which was confirmed by its absorbance and fluorescence spectra and retention time on high-performance liquid chromatography. The PLP-OADS oxime was displaced from the enzyme by the addition of PLP as evidenced by the restoration of complete enzyme activity as well as by the spectral properties. The unique feature of the mechanism proposed for the interaction of OADS with sheep liver SHMT was the formation of PLP as an intermediate.
 
Date 1989
 
Type Article
PeerReviewed
 
Format pdf
 
Language en
 
Identifier http://ir.cftri.com/2695/1/Biochemistry%2C%20Vol.%2028%2C%20No.%2025%2C%201989.pdf
Baskaran, N. and Prakash, V. and Appu Rao, A. G. and Radhakrishnan, A. N. and Savithri, H. S. and Appaji Rao, N. (1989) Mechanism of interaction of O-amino-D-serine with sheep liver serine hydroxymethyltransferase. Biochemistry, 28. pp. 9607-9612.