Kinetic mechanism of the interaction of D-cycloserine with serine hydroxymethyltransferase.
IR@CSIR-CFTRI
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http://ir.cftri.com/4532/
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Title |
Kinetic mechanism of the interaction of D-cycloserine with serine hydroxymethyltransferase.
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Creator |
Manohar, R.
Appu Rao, A. G. Appaji Rao, N. |
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Subject |
07 Enzyme Biochemistry
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Description |
The kinetic mechanism for the interaction of D-cycloserine with serine hydroxymethyltransferase (EC 2.1.2.1) from sheep liver was established by measuring changes in the activity, absorbance, and circular dichoism (CD) of the enzyme. The irreversible inhibition of the enzyme was characterized by three detectable steps: an initial rapid step followed by two successive steps with rate constants of 5.4 X s-l and 1.4 X lo4 s-l. The first step was distinguished by a rapid disappearance of the enzyme absorbance peak at 425 nm, a decrease in the enzyme activity to 25% of the uninhibited velocity, and a lowering of the CD intensity at 432 nm to about 65% of the original value. The second step of the interaction was accompanied by a complete loss of enzyme activity and a marginal increase in the CD intensity at 432 nm. The final step resulted in the complete loss of the enzyme absorbance at 425 nm and of the CD band at 432 nm. The products of the reaction were identified as (a) apoenzyme by absorbance measurements, CD spectra, and reconstitution with pyridoxal 5’-phosphate and (b) a pyridoxal 5’-phosphate-~- cycloserine Schiff s base complex identified by its fluorescence and absorbance spectra. The Schiff base complex was expelled from the enzyme active site in the final step of the reaction. The proposed mechanism, which is different from those operative in other pyridoxal phosphate dependent enzymes, probably accounts for the selective inhibition of serine hydroxymethyltransferase by the drug in vivo. |
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Date |
1984
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Type |
Article
PeerReviewed |
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Format |
pdf
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Language |
en
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Identifier |
http://ir.cftri.com/4532/1/Biochemistry%201984%2C%2023%2C%2041%2016-4122.pdf
Manohar, R. and Appu Rao, A. G. and Appaji Rao, N. (1984) Kinetic mechanism of the interaction of D-cycloserine with serine hydroxymethyltransferase. Biochemistry, 23. pp. 4116-4122. |
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