Purification and partial amino acid sequence analysis of human erythrocyte acetylcholinesterase

Vijay Chhajlani; Dwight Derr; Betty Earles; Eli Schmell; Thomas August

doi:10.1016/0014-5793(89)81352-3

Purification and partial amino acid sequence analysis of human erythrocyte acetylcholinesterase

Vijay Chhajlani, Dwight Derr, Betty Earles, Eli Schmell, Thomas August

Research output: Contribution to journal › Article › peer-review

15 Scopus citations

Abstract

A single step immunoaffinity purification procedure for human erythrocyte acetylcholinesterase is described which permitted the isolation of milligram quantities of enzyme from 10 U of erythrocytes, with 113 000-fold purification and a yield of about 22%. In SDS-PAGE analysis, the enzyme corresponds to a disulfide linked dimer of 140 kDa which is converted to a 70 kDa monomer upon disulfide reduction. The tryptic peptides generated from purified enzyme were separated by reverse-phase HPLC. Five of these peptides were analysed to determine the amino acid sequences. The obtained sequences showed no homology to the already known amino acid sequences for human serum and brain butyryl-cholinesterase and Torpedo californica acetylcholinesterase.

Original language	English (US)
Pages (from-to)	279-282
Number of pages	4
Journal	FEBS Letters
Volume	247
Issue number	2
DOIs	https://doi.org/10.1016/0014-5793(89)81352-3
State	Published - Apr 24 1989
Externally published	Yes

Keywords

Acetylcholinesterase
Amino acid sequence
Immunoaffinity chromatography

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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10.1016/0014-5793(89)81352-3

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abstract = "A single step immunoaffinity purification procedure for human erythrocyte acetylcholinesterase is described which permitted the isolation of milligram quantities of enzyme from 10 U of erythrocytes, with 113 000-fold purification and a yield of about 22%. In SDS-PAGE analysis, the enzyme corresponds to a disulfide linked dimer of 140 kDa which is converted to a 70 kDa monomer upon disulfide reduction. The tryptic peptides generated from purified enzyme were separated by reverse-phase HPLC. Five of these peptides were analysed to determine the amino acid sequences. The obtained sequences showed no homology to the already known amino acid sequences for human serum and brain butyryl-cholinesterase and Torpedo californica acetylcholinesterase.",

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T1 - Purification and partial amino acid sequence analysis of human erythrocyte acetylcholinesterase

AU - Chhajlani, Vijay

AU - Derr, Dwight

AU - Earles, Betty

AU - Schmell, Eli

AU - August, Thomas

PY - 1989/4/24

Y1 - 1989/4/24

N2 - A single step immunoaffinity purification procedure for human erythrocyte acetylcholinesterase is described which permitted the isolation of milligram quantities of enzyme from 10 U of erythrocytes, with 113 000-fold purification and a yield of about 22%. In SDS-PAGE analysis, the enzyme corresponds to a disulfide linked dimer of 140 kDa which is converted to a 70 kDa monomer upon disulfide reduction. The tryptic peptides generated from purified enzyme were separated by reverse-phase HPLC. Five of these peptides were analysed to determine the amino acid sequences. The obtained sequences showed no homology to the already known amino acid sequences for human serum and brain butyryl-cholinesterase and Torpedo californica acetylcholinesterase.

AB - A single step immunoaffinity purification procedure for human erythrocyte acetylcholinesterase is described which permitted the isolation of milligram quantities of enzyme from 10 U of erythrocytes, with 113 000-fold purification and a yield of about 22%. In SDS-PAGE analysis, the enzyme corresponds to a disulfide linked dimer of 140 kDa which is converted to a 70 kDa monomer upon disulfide reduction. The tryptic peptides generated from purified enzyme were separated by reverse-phase HPLC. Five of these peptides were analysed to determine the amino acid sequences. The obtained sequences showed no homology to the already known amino acid sequences for human serum and brain butyryl-cholinesterase and Torpedo californica acetylcholinesterase.

KW - Acetylcholinesterase

KW - Amino acid sequence

KW - Immunoaffinity chromatography

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Purification and partial amino acid sequence analysis of human erythrocyte acetylcholinesterase

Abstract

Keywords

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