TY - JOUR
T1 - On the relationship between nucleotide hydrolysis and microtubule assembly
T2 - Studies with a GTP-regenerating system
AU - Schilstra, M.
AU - Martin, S.R.
AU - Bayley, P.M.
N1 - Original article can be found at: http://www.sciencedirect.com/science/journal/0006291X Copyright Elsevier Inc. [Full text of this article is not available in the UHRA]
PY - 1987
Y1 - 1987
N2 - The assembly of pure tubulin dimer has been studied in two buffer systems (containing low and high glycerol/Mg), using a regeneration system protocol to assess the amount of GDP-tubulin in the assembling polymer. For both assembly systems studied, the GDP content is effectively stoichiometric with tubulin throughout assembly. This indicates a high degree of coupling between assembly and GTP-hydrolysis, giving a hydrolysis rate at least 10-fold faster than previously deduced. The steady state GTP hydrolysis rate is quantitatively consistent with this finding. We conclude that the extent of any GTP-tubulin cap is below the detectable limit, both during elongation and at steady state. MT-protein; microtubule protein; MAPs; microtubule associated proteins; PC-tubulin; tubulin dimer from phosphocellulose chromatography; GTP- and GDP-tubulin denotes the nucleotide at the exchangeable nucleotide binding site (E-site)RS; enzymic GTP-regenerating system.
AB - The assembly of pure tubulin dimer has been studied in two buffer systems (containing low and high glycerol/Mg), using a regeneration system protocol to assess the amount of GDP-tubulin in the assembling polymer. For both assembly systems studied, the GDP content is effectively stoichiometric with tubulin throughout assembly. This indicates a high degree of coupling between assembly and GTP-hydrolysis, giving a hydrolysis rate at least 10-fold faster than previously deduced. The steady state GTP hydrolysis rate is quantitatively consistent with this finding. We conclude that the extent of any GTP-tubulin cap is below the detectable limit, both during elongation and at steady state. MT-protein; microtubule protein; MAPs; microtubule associated proteins; PC-tubulin; tubulin dimer from phosphocellulose chromatography; GTP- and GDP-tubulin denotes the nucleotide at the exchangeable nucleotide binding site (E-site)RS; enzymic GTP-regenerating system.
U2 - 10.1016/0006-291X(87)90971-5
DO - 10.1016/0006-291X(87)90971-5
M3 - Article
SN - 0006-291X
VL - 147
SP - 588
EP - 595
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 2
ER -