Association of phospholamban with a cGMP kinase signaling complex.
The cGMP kinase signaling complex identified previously in tracheal smooth muscle membranes contains a number of cGMP kinase substrates termed G0 through G4. G0, G1, and G2 were identified as IP(3) receptor I (IP(3)RI), IRAG, and cGMP kinase I. Sequencing of purified G3 and G4 showed that these proteins were proteolytic cleavage products of IRAG. However, the purified cGMP kinase signaling complex contained following additional proteins: alpha-actin, calponin H1, and phospholamban (PLB) as verified by MALDI-TOF as well as MS/MS sequencing and immune detection. The complex of these six proteins was immune precipitated by antibodies to each protein. The proteins were phosphorylated by the endogenous cGMP kinase I with the exception of alpha-actin and calponin H1. The complex did not contain the Ca(2+)-ATPase SERCA II. PLB, IP(3)RI, and cGMP kinase Ibeta were co-immune precipitated after expression in COS-7 cells. These results suggest that PLB may have additional functions to regulate the activity of SERCA II.
Koller
Angela
A
Institut für Pharmakologie und Toxikologie, TU München, Biedersteiner Strasse 29, 80802 Munich, Germany. koller@ipt.med.tu-muenchen.de
Schlossmann
Jens
J
Ashman
Keith
K
Uttenweiler-Joseph
Sandrine
S
Ruth
Peter
P
Hofmann
Franz
F
D000199:Actins / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D019556:COS Cells; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000737:chemistry / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry / Q000378:metabolism; D002417:Cattle; D062965:Cyclic GMP-Dependent Protein Kinase Type I; D017869:Cyclic GMP-Dependent Protein Kinases / Q000737:chemistry / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000378:metabolism; D066298:In Vitro Techniques; D047908:Intracellular Signaling Peptides and Proteins; D046911:Macromolecular Substances; D008565:Membrane Proteins; D051379:Mice; D008840:Microfilament Proteins; D009130:Muscle, Smooth / Q000378:metabolism; D010750:Phosphoproteins / Q000737:chemistry / Q000378:metabolism; D010766:Phosphorylation; D011233:Precipitin Tests; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D015398:Signal Transduction; D013379:Substrate Specificity; D000096985:Calponins; D000099211:Phospholamban
D000199:Actins; D002135:Calcium-Binding Proteins; D000252:Calcium-Transporting ATPases; D002352:Carrier Proteins; D062965:Cyclic GMP-Dependent Protein Kinase Type I; D017869:Cyclic GMP-Dependent Protein Kinases; D004268:DNA-Binding Proteins; D047908:Intracellular Signaling Peptides and Proteins; D046911:Macromolecular Substances; D008565:Membrane Proteins; D008840:Microfilament Proteins; D010750:Phosphoproteins; D011994:Recombinant Proteins; D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases; D000096985:Calponins; D000099211:Phospholamban; C112986:AIF1 protein, human; C485784:IRAG1 protein, human; C406072:MRVI1 protein, Bos taurus; C485785:Mrvi1 protein, mouse; C015482:protein kinase modulator; C569718:PRKG1 protein, human; C569720:Prkg1 protein, mouse