Evidence for Rap1 in vascular smooth muscle cells

FEBS Letters

Enouf, J.¹  Fontenay, M.¹  de Gunzburg, J.²  Bredoux, R.¹  Corvazier, E.¹  Quarck, R.¹  Tobelem, G.¹  Bryckaert, M.¹  Magmer, C.¹ 

[1] INSERM U. 348, Hôpital Lariboisière, 6 rue Guy Patin, 75010 Paris, France;INSERM U. 248, Faculté de Médecine Lariboisière-Saint Louis, 10 avenue de Verdun, 75010 Paris, France

关键词: Platelet-derived growth factor;    Rap1;    Vascular smooth muscle cell;    rap;    Ras-proximate;    PDGF;    platelet-derived growth factor;    SMC;    smooth muscle cells;    PKA;    cAMP-dependent protein kinase;    C. Sub.;    catalytic subunit of PKA;    HEPES;    4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid;    EGTA;    [ethylene bis (oxyethylenenitrilo)]tetraacetic acid;    DTT;    dithiothreitol;    SBTI;    soybean trypsin inhibitor;    SDS;    sodium dodecyl sulfate;    PAGE;    polyacrylamide gel electrophoresis;    RT-PCR;    reverse transcription-PCR;    SERCA;    sarco/endoplasmic reticulum Ca2+ ATPase;    MMLV;    murine Moloney leukemia virus;    SSC;    sodium salt citrate;    GAP;    GTPase activating protein;   

DOI  :  10.1016/0014-5793(94)80492-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The effect of platelet-derived growth factor (PDGF) on Rap1 expression was investigated in rat vascular smooth muscle cells (SMC). First, evidence for Rap1 proteins was shown by their: (i) detection in membranes using a specific anti-Rap1 antibody, (ii) typical shift in electrophoretic mobility as a consequence of reduction, and (iii) cAMP-induced phosphorylation and immunoprecipitation. Then, the mitogenic activity of 10 PDGF AA and BB for 48 h, resulting in a 2- and 5-fold increase in [³H]thymidine incorporation, was correlated with that of total Rap1 protein expression which was found to be 99% ± 36% and 260% ± 70%, respectively. Further time-course studies established that this up-regulation of Rap1 proteins was only observed after 48 h of PDGF BB treatment. Lastly, comparative RT-PCR of both rap1a and rap1b mRNAs showed that PDGF BB also up-regulated the rap1a mRNA species, which was 1.5-fold increased in contrast with the rap1b mRNA species. It is concluded that the PDGF BB-induced SMC proliferation is associated with an up-regulation of Rap1a protein.

【 授权许可】

Unknown