The Purification of Choline Acetyltransferase of Squid-head Ganglia

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1973 Dec 1

PMID 4521199

Citations 9

Authors

S S Husain

H G MAUTNER

Affiliations

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Abstract

Choline acetyltransferase (EC 2.3.1.6) isolated from the head ganglia of squid could be purified by use of mercurial-Sepharose columns as well as Sepharose columns to which the enzyme inhibitor p-(m-bromophenyl)vinyl pyridinium had been attached. These columns, in conjunction with 30-55% ammonium sulfate precipitation, 40-30% ammonium sulfate extraction, chromatography on sulfopropyl-Sephadex and on cellulose phosphate and hydroxylapatite columns, led to the isolation of three factions of choline acetyltransferase ranging in activity from 1000 to 4000 mumole/mg of protein/per hr. Polyacrylamide gel electrophoresis suggests that two of these fractions are homogeneous. The squid choline acetyltransferase is different from the mammalian-brain enzymes in having a larger molecular weight under the conditions used and in being relatively poorly inhibited by styryl pyridinium compounds.

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References

Cuatrecasas P . Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads. J Biol Chem. 1970; 245(12):3059-65. View

Glover V, Potter L . Purification and properties of choline acetyltransferase from ox brain striate nuclei. J Neurochem. 1971; 18(4):571-80. DOI: 10.1111/j.1471-4159.1971.tb11987.x. View

Morris D, Maneckjee A, HEBB C . The kinetic properties of human placental choline acetyltransferase. Biochem J. 1971; 125(3):857-63. PMC: 1178191. DOI: 10.1042/bj1250857. View

Nachmansohn D, Wilson I . The enzymic hydrolysis and synthesis of acetylcholine. Adv Enzymol Relat Subj Biochem. 1951; 12:259-339. DOI: 10.1002/9780470122570.ch5. View

Allfrey V . A method for the purification of histone fraction F3 by affinity chromatography. Arch Biochem Biophys. 1973; 154(1):185-91. DOI: 10.1016/0003-9861(73)90047-7. View

Cuatrecasas P . Affinity chromatography. Annu Rev Biochem. 1971; 40:259-78. DOI: 10.1146/annurev.bi.40.070171.001355. View

Davis B . DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964; 121:404-27. DOI: 10.1111/j.1749-6632.1964.tb14213.x. View

SLUYTERMAN L, Wijdenes J . An agarose mercurial column for the separation of mercaptopapain and nonmercaptopapain. Biochim Biophys Acta. 1970; 200(3):593-5. DOI: 10.1016/0005-2795(70)90122-4. View

Schrier B, Shuster L . A simplified radiochemical assay for choline acetyltransferase. J Neurochem. 1967; 14(10):977-85. DOI: 10.1111/j.1471-4159.1967.tb09509.x. View

10.

Prince A . Properties of choline acetyltransferase isolated from squid ganglia. Proc Natl Acad Sci U S A. 1967; 57(4):1117-22. PMC: 224663. DOI: 10.1073/pnas.57.4.1117. View

11.

SASTRY B, Henderson G . Kinetic mechanisms of human placental choline acetyltransferase. Biochem Pharmacol. 1972; 21(6):787-802. DOI: 10.1016/0006-2952(72)90122-0. View

12.

Dudai Y, Silman I, Shinitzky M, Blumberg S . Purification by affinity chromatography of the molecular forms of acetylcholinesterase present in fresh electric-organ tissue of electric eel. Proc Natl Acad Sci U S A. 1972; 69(9):2400-3. PMC: 426949. DOI: 10.1073/pnas.69.9.2400. View

13.

Chao L, WOLFGRAM F . Purification and some properties of choline acetyltransferase (EC 2.3.1.6) from bovine brain. J Neurochem. 1973; 20(4):1075-81. DOI: 10.1111/j.1471-4159.1973.tb00078.x. View

14.

Leuzinger W, Baker A, Cauvin E . Acetylcholinesterase. II. Crystallization, absorption spectra, isoionic point. Proc Natl Acad Sci U S A. 1968; 59(2):620-3. PMC: 224717. DOI: 10.1073/pnas.59.2.620. View

15.

Prince A, Hide E . Activation of choline acetyltransferase by salts. Nat New Biol. 1971; 234(50):222-3. DOI: 10.1038/newbio234222a0. View

16.

Andrews P . Estimation of molecular size and molecular weights of biological compounds by gel filtration. Methods Biochem Anal. 1970; 18:1-53. View

17.

Nachmansohn D . Metabolism and function of the nerve cell. Harvey Lect. 1953; 49:57-99. View

18.

Fonnum F . Multiple forms of choline acetyltransferase in several species demonstrated by isoelectric focusing. Biochem J. 1972; 127(1):229-36. PMC: 1178577. DOI: 10.1042/bj1270229. View

19.

LOWRY O, ROSEBROUGH N, FARR A, RANDALL R . Protein measurement with the Folin phenol reagent. J Biol Chem. 1951; 193(1):265-75. View

20.

Potter L, Glover V, SAELENS J . Choline acetyltransferase from rat brain. J Biol Chem. 1968; 243(14):3864-70. View