HSP27 in signal transduction and association with contractile proteins in smooth muscle cells.
Sustained smooth muscle contraction is mediated by protein kinase C (PKC) through a signal transduction cascade leading to contraction. Heat-shock protein 27 (HSP27) appears to be the link between these two major events, i.e., signal transduction and sustained smooth muscle contraction. We have investigated the involvement of HSP27 in signal transduction and HSP27 association with contractile proteins (e.g., actin, myosin, tropomyosin, and caldesmon) resulting in sustained smooth muscle contraction. We have carried out confocal microscopy to investigate the cellular reorganization and colocalization of proteins and immunoprecipitation of HSP27 with actin, myosin, tropomyosin, and caldesmon as detected by sequential immunoblotting. Our results indicate that 1) translocation of Raf-1 to the membrane when stimulated with ceramide is inhibited by vasoactive intestinal peptide (VIP), a relaxant neuropeptide; 2) PKC-alpha and mitogen-activated protein kinase translocate and colocalize on the membrane in response to ceramide, and PKC-alpha translocation is inhibited by VIP; 3) HSP27 colocalizes with actin when contraction occurs; and 4) HSP27 immunoprecipitates with actin and with the contractile proteins myosin, tropomyosin, and caldesmon. We propose a model in which HSP27 is involved in sustained smooth muscle contraction and modulates the interaction of actin, myosin, tropomyosin, and caldesmon.
Ibitayo
A I
AI
Department of Pediatrics, University of Michigan Medical Center, Ann Arbor, Michigan 48109, USA.
Sladick
J
J
Tuteja
S
S
Louis-Jacques
O
O
Yamada
H
H
Groblewski
G
G
Welsh
M
M
Bitar
K N
KN
D000199:Actins / Q000378:metabolism; D000818:Animals; D001381:Azepines / Q000494:pharmacology; D001692:Biological Transport; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D002148:Calmodulin-Binding Proteins / Q000378:metabolism; D002518:Ceramides / Q000494:pharmacology; D003106:Colon / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D003285:Contractile Proteins / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000378:metabolism / Q000502:physiology*; D007527:Isoenzymes / Q000378:metabolism; D009119:Muscle Contraction / Q000187:drug effects / Q000502:physiology; D009130:Muscle, Smooth / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D009218:Myosins / Q000378:metabolism; D009281:Naphthalenes / Q000494:pharmacology; D011233:Precipitin Tests; D011493:Protein Kinase C / Q000378:metabolism; D051571:Protein Kinase C-alpha; D019908:Proto-Oncogene Proteins c-raf / Q000378:metabolism; D011817:Rabbits; D015398:Signal Transduction / Q000502:physiology*; D014018:Tissue Distribution; D014335:Tropomyosin / Q000378:metabolism
D000199:Actins; D001381:Azepines; D002148:Calmodulin-Binding Proteins; D002518:Ceramides; D003285:Contractile Proteins; D006360:Heat-Shock Proteins; D007527:Isoenzymes; D009281:Naphthalenes; D014335:Tropomyosin; C070571:ML 7; D019908:Proto-Oncogene Proteins c-raf; D011493:Protein Kinase C; D051571:Protein Kinase C-alpha; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D009218:Myosins