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PMID: 191810 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Stoichiometry of GTP hydrolysis and tubulin polymerization.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 74 ·No. 2 ·1977-02-00 ·Pages 462-6

Maccioni R, Seeds NW

Abstract

Microtubule formation from lamb brain tubulin isolated by affinity chromatography and freed of exchangeable nucleotide requires GTP for maximal rate and extent of polymerization. The nucleotide analogs guanylylmethylenediphosphate and guanylylimidodiphosphate fail to replace GTP; in addition, neither the presence of microtubule associated proteins nor 5 M glycerol relieves the GTP requirement. The relation of GTP concentration and microtubule formation shows an association constant K = 1 X 10(4) M-1; furthermore, GDP and guanylylimidodiphosphate are competitive inhibitors of GTP for polymerization. Using a rapid filter assay for microtubule formation that allows the quantitative analysis of early polymerization kinetics and correcting for GTP hydrolysis uncoupled from tubulin polymerization, a stoichiometry of two molecules of GTP hydrolyzed per mole of tubulin dimer incorporated into microtubules has been found.

MeSH Terms
Animals Brain/metabolism Chromatography, Affinity Glycoproteins/metabolism Guanosine Triphosphate/analogs & derivatives,metabolism Kinetics Macromolecular Substances Microtubules/metabolism Phosphoric Diester Hydrolases/metabolism Protein Binding Sheep Tubulin/isolation & purification,metabolism
Chemicals
Glycoproteins Macromolecular Substances Tubulin Guanosine Triphosphate Phosphoric Diester Hydrolases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Maccioni R
Seeds N W
References (19)
19 references, click to expand
  1. Properties of tubulin prepared by affinity chromatography.
    Ann N Y Acad Sci. 1975 Jun 30;253:260-71 PMID: 1056745
  2. Tubulin nucleotide reactions and their role in microtubule assembly and dissociation.
    Ann N Y Acad Sci. 1975 Jun 30;253:562-72 PMID: 1096723
  3. Microtubular protein reaction with nucleotides.
    Biochem Biophys Res Commun. 1976 Jan 12;68(1):127-35 PMID: 1247450
  4. Effect of guanine nucleotides on the assembly of brain microtubles: ability of 5'-guanylyl imidodiphosphate to replace GTB in promoting the polymerization of microtubules in vitro.
    Biochem Biophys Res Commun. 1976 Mar 22;69(2):369-76 PMID: 131550
  5. Protein measurement with the Folin phenol reagent.
    J Biol Chem. 1951 Nov;193(1):265-75 PMID: 14907713
  6. Ionic and nucleotide requirements for microtubule polymerization in vitro.
    Biochemistry. 1975 Jul;14(13):2996-3005 PMID: 238580
  7. Purification of tubulin from bovine brain and its interaction with guanine nucleotides.
    J Biochem. 1975 Mar;77(3):647-58 PMID: 238963
  8. Comparison of guanosine triphosphate split and polypeptide synthesis with a purified E. coli system.
    Proc Natl Acad Sci U S A. 1966 Jan;55(1):212-9 PMID: 4287350
  9. Adenylyl imidodiphosphate, an adenosine triphosphate analog containing a P--N--P linkage.
    Biochemistry. 1971 Jun 22;10(13):2484-9 PMID: 4326768
  10. Turbidimetric studies of the in vitro assembly and disassembly of porcine neurotubules.
    J Mol Biol. 1974 Nov 15;89(4):737-55 PMID: 4475698
  11. Microtubule assembly in the absence of added nucleotides.
    Proc Natl Acad Sci U S A. 1973 Mar;70(3):765-8 PMID: 4514990
  12. Microtubule formation in vitro in solutions containing low calcium concentrations.
    Science. 1972 Sep 22;177(4054):1104-5 PMID: 4626639
  13. Isolation of brain tubulin by affinity chromatography.
    Biochem Biophys Res Commun. 1973 Jun 8;52(3):752-8 PMID: 4710561
  14. Microtubule assembly in vitro.
    Fed Proc. 1974 Feb;33(2):167-74 PMID: 4811890
  15. A circular dichroism study of microtubule protein.
    Biochemistry. 1972 Apr 25;11(9):1554-61 PMID: 5028102
  16. Nucleated assembly of microtubules in porcine brain extracts.
    Science. 1972 Sep 29;177(4055):1196-7 PMID: 5057625
  17. The colchicine-binding protein of mammalian brain and its relation to microtubules.
    Biochemistry. 1968 Dec;7(12):4466-79 PMID: 5700666
  18. Guanine nucleotide associated with the protein of the outer fibers of flagella and cilia.
    Science. 1967 Jun 23;156(3782):1606-8 PMID: 6067301
  19. Biochemistry and physiology of microtubules.
    Annu Rev Biochem. 1976;45:699-720 PMID: 786160
Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.74.2.462
Published
1977-02-00
Pages
462-6
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC392309
Subset
IM
External Links
PubMed source DOI: 10.1073/pnas.74.2.462 PMC Full Text View journal details
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