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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
39
pubmed:dateCreated
1994-11-8
pubmed:abstractText
The exchangeable nucleotide-binding site of tubulin has been studied using diastereoisomers A (Sp) and B (Rp) of guanosine 5'-O-(1-thiotriphosphate) (GTP alpha S) in which the phosphorus atom to which sulfur is attached is chiral. GTP alpha S(A) (10 microM) nucleated assembly of purified tubulin (20 microM) into microtubules in buffer containing 0.1 M 2-(N-morpholino)ethanesulfonic acid with 3 mM Mg2+ and 1 mM EGTA, pH 6.6 at 37 degrees C. With 0.2 mM GTP alpha S(A), the critical concentration (Cc; minimum protein concentration required for assembly) was 8 microM tubulin. Neither 0.2 mM GTP nor GTP alpha S(B) promoted microtubule assembly in buffer with 0.5-6.75 mM Mg2+ and 20-70 microM tubulin. The Cc values for GTP alpha S-(A)-induced assembly of tubulin in buffer with 30% glycerol and of microtubule protein (tubulin and microtubule-associated proteins) in buffer were lower than for GTP. GTP alpha S(A)-induced microtubules were more stable to the cold and to Ca2+. GTP alpha S(A) and GTP but not GTP alpha S(B) bound tightly to tubulin at 4 degrees C. Although GTP alpha S(B) did not nucleate assembly, it did bind to tubulin since it was incorporated into the growing microtubule. Both isomers were hydrolyzed in the microtubules. These studies show that GTP alpha S(A) promotes tubulin assembly better than GTP and GTP alpha S(B) and that there is stereoselectivity at the alpha-phosphate binding region of tubulin. The stereoselectivity may be due to different MgGTP alpha S(A) and -(B) interactions with tubulin.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Oct
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
4
pubmed:volume
33
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
11884-90
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed:year
1994
pubmed:articleTitle
Interaction of tubulin with guanosine 5'-O-(1-thiotriphosphate) diastereoisomers: specificity of the alpha-phosphate binding region.
pubmed:affiliation
Department of Psychiatric Medicine, University of Virginia School of Medicine, Charlottesville 22908.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.