PMID- 11015208
OWN - NLM
STAT- MEDLINE
DCOM- 20001121
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 39
IP  - 40
DP  - 2000 Oct 10
TI  - Stathmin slows down guanosine diphosphate dissociation from tubulin in a 
      phosphorylation-controlled fashion.
PG  - 12295-302
AB  - Stathmin is an important protein that interacts with tubulin and regulates 
      microtubule dynamics in a phosphorylation-controlled fashion. Here we show that 
      the dissociation of guanosine 5'-diphosphate (GDP) from beta-tubulin is slowed 
      20-fold in the (tubulin)(2)-stathmin ternary complex (T(2)S). The kinetics of GDP 
      or guanosine 5'-triphosphate (GTP) dissociation from tubulin have been monitored 
      by the change in tryptophan fluorescence of tubulin upon exchanging 
      2-amino-6-mercapto-9-beta-ribofuranosylpurine 5'-diphosphate (S6-GDP) for 
      tubulin-bound guanine nucleotide. At molar ratios of stathmin to tubulin lower 
      than 0.5, biphasic kinetics were observed, indicating that the dynamics of the 
      complex is extremely slow, consistent with its high stability. The method was 
      used to characterize the effects of phosphorylation of stathmin on its 
      interaction with tubulin. The serine-to-glutamate substitution of all four 
      phosphorylatable serines of stathmin (4E-stathmin) weakens the stability of the 
      T(2)S complex by about 2 orders of magnitude. The phosphorylation of serines 16 
      and 63 in stathmin has a more severe effect and weakens the stability of T(2)S 
      10(4)-fold. The rate of GDP dissociation is lowered only 7-fold and 4-fold in the 
      complexes of tubulin with 4E-stathmin and diphosphostathmin, respectively. 
      Sedimentation velocity studies support the conclusions of nucleotide exchange 
      data and show that the T(2)S complexes formed between tubulin and 4E-stathmin or 
      diphosphostathmin are less compact than the highly stable T(2)S complex. The 
      correlation between the effect of phosphorylation of stathmin on the stability of 
      T(2)S complex measured in vitro and on the function of stathmin in vivo is 
      discussed.
FAU - Amayed, P
AU  - Amayed P
AD  - Dynamique du Cytosquelette, Laboratoire d'Enzymologie et Biochimie Structurale 
      Centre National de la Recherche Scientifique, 91198 Gif-sur-Yvette, France.
FAU - Carlier, M F
AU  - Carlier MF
FAU - Pantaloni, D
AU  - Pantaloni D
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Guanine Nucleotide Exchange Factors)
RN  - 0 (Macromolecular Substances)
RN  - 0 (Microtubule Proteins)
RN  - 0 (Phosphoproteins)
RN  - 0 (Stathmin)
RN  - 0 (Tubulin)
RN  - 146-91-8 (Guanosine Diphosphate)
RN  - 86-01-1 (Guanosine Triphosphate)
RN  - 8DUH1N11BX (Tryptophan)
SB  - IM
MH  - Animals
MH  - Cattle
MH  - Guanine Nucleotide Exchange Factors/metabolism/*physiology
MH  - Guanosine Diphosphate/*metabolism
MH  - Guanosine Triphosphate/metabolism
MH  - Kinetics
MH  - Macromolecular Substances
MH  - *Microtubule Proteins
MH  - Phosphoproteins/metabolism/*physiology
MH  - Phosphorylation
MH  - Spectrometry, Fluorescence
MH  - Stathmin
MH  - Tryptophan
MH  - Tubulin/*metabolism
MH  - Ultracentrifugation
EDAT- 2000/10/04 11:00
MHDA- 2001/02/28 10:01
CRDT- 2000/10/04 11:00
PHST- 2000/10/04 11:00 [pubmed]
PHST- 2001/02/28 10:01 [medline]
PHST- 2000/10/04 11:00 [entrez]
AID - bi000279w [pii]
AID - 10.1021/bi000279w [doi]
PST - ppublish
SO  - Biochemistry. 2000 Oct 10;39(40):12295-302. doi: 10.1021/bi000279w.