Purified human serum butyrylcholinesterase after treatment with either of the metal chelators EDTA or NaCN was able to bind to a Zn2+-chelate–Sepharose affinity column and was eluted from the column by EDTA or imidazole. Prior EDTA treatment of the enzyme was essential for binding to this affinity column. The enzyme could be labelled with 65Zn2+ after EDTA treatment of the enzyme. Diethylpyrocarbonate modification of histidine residues in the EDTA-treated enzyme resulted in the abolition of both binding to the Zn2+-chelate–Sepharose column and labelling by 65Zn2+. Stoicheiometry of 65Zn2+ binding indicated approximately 0.85 mol of Zn2+/mol of subunit of the EDTA-treated enzyme. EDTA or NaCN treatment resulted in the loss of thermal stability of the enzyme at 37 °C which could not be reversed by Zn2+. Whereas the cholinesterase activity of butyrylcholinesterase was not affected by EDTA, there was significant loss of its carboxypeptidase activity in the presence of EDTA, and the loss could be reversed by added ZnCl2. These results suggest the presence of a Zn2+-binding site on human serum butyrylcholinesterase and the involvement of histidine residues in the metal binding. The presence in human serum butyrylcholinesterase of a sequence HXXE…H found in many known Zn2+-containing enzymes supports these findings.
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April 1996
Research Article|
April 01 1996
Evidence for a Zn2+-binding site in human serum butyrylcholinesterase
C. D. BHANUMATHY;
C. D. BHANUMATHY
* Neurochemistry Laboratory, Department of Neurological Sciences, Christian Medical College and Hospital, Vellore 632 004, India
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A. S. BALASUBRAMANIAN
A. S. BALASUBRAMANIAN
* Neurochemistry Laboratory, Department of Neurological Sciences, Christian Medical College and Hospital, Vellore 632 004, India
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Biochem J (1996) 315 (1): 127–131.
Article history
Received:
August 09 1995
Revision Received:
November 17 1995
Accepted:
December 01 1995
Citation
C. D. BHANUMATHY, A. S. BALASUBRAMANIAN; Evidence for a Zn2+-binding site in human serum butyrylcholinesterase. Biochem J 1 April 1996; 315 (1): 127–131. doi: https://doi.org/10.1042/bj3150127
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