Purification of tubulin from bovine brain and its interaction with guanine nucleotides.

Purification of tubulin from bovine brain and its interaction with guanine nucleotides. Research Abstract Details 

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Purification of tubulin from bovine brain and its interaction with guanine nucleotides. Abstract Text:

T Arai,Y Ihara,K Arai,Y Kaziro,

A rapid and sensitive assay for [3H]GTP binding activity of tubulin has been developed. This assay method is based on the quantitative retention of [3H]GTP. Tubulin complex on a nitrocellulose membrane filter. It was also found that bovine brain tubulin is markedly stablized by glycerol and GTP against denaturation. A large-scale purification of bovine brain tubulin was achieved using the new assay procedure and by the inclusion of glycerol and GTP in a buffer solution used for column chromatograph. The purified tubulin could be stored at -80degrees in the presence of glycerol and GTP for at least a year without any apprecialbe loss of [3H]GTP- and [3H]colchicine binding activities. The interaction of tubulin with guanine nucleotides was also studied using the nitorcellulose membrane filter procedure. It was found that the binding of [3H]GTP to tubulin with an empty exchangeable site proceeded promptly within k sec while the exchange of [3H]GTP- with a GTP-tubulin complex in which the exchangeable site had been occupied with unlabeled GTP occured more slowly. The dissociation constants for GTP and GDP at the exchangeable site of tubulin were determined as 0.5 times 10-6M and 1.9 times 10-6M, respectively. 5'-Guanylylimidodiphosphate could interact, although less strongly, with tubulin at this site, whereas the interaction of other nucleoside triphosphates includint ATP, CTP, UTP, and 5'-guanylyl methylenediphosphonate was very weak, if it occured at all. The presence of Mg2+ and a free sulfhydryl group was found to be essential for binding of [3H]GTP to tubulin. Ca2+ was found to replace Mg2+ in this binding reaction.

Purification of tubulin from bovine brain and its interaction with guanine nucleotides. Publishing Authors By Initials

T Arai,Y Ihara,K Arai,Y Kaziro,

For similar macromolecular substances: polymers: biopolymers: microtubule proteins: tubulin research abstracts see: macromolecular substances: polymers: biopolymers: microtubule proteins: tubulin research

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Purification of tubulin from bovine brain and its interaction with guanine nucleotides. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Journal of biochemistry

VOLUME: 77

Page Numbers: 647-58

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: Mar

YEAR: 1975

Purification of tubulin from bovine brain and its interaction with guanine nucleotides. Information

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LANGUAGE: eng

NlmUniqueID: 376600

Purification of tubulin from bovine brain and its interaction with guanine nucleotides. Keywords Mesh Terms:

KEYWORDS: Tubulin

MESH TERMS: metabolism

Chemical & Substance for Abstract: Purification of tubulin from bovine brain and its interaction with guanine nucleotides. Information

Substance Name: Guanosine Triphosphate

Registry Number: 86-01-1

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Country: JAPAN

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MEDLINETA: J Biochem

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