p23/Tmp21 differentially targets the Rac-GAP beta2-chimaerin and protein kinase C via their C1 domains.
The C1 domains in protein kinase C (PKC) isozymes and other signaling molecules are responsible for binding the lipid second messenger diacylglycerol and phorbol esters, and for mediating translocation to membranes. Previous studies revealed that the C1 domain in alpha- and beta-chimaerins, diacylglycerol-regulated Rac-GAPs, interacts with the endoplasmic reticulum/Golgi protein p23/Tmp21. Here, we found that p23/Tmp21 acts as a C1 domain-docking protein that mediates perinuclear translocation of beta2-chimaerin. Glu227 and Leu248 in the beta2-chimaerin C1 domain are crucial for binding p23/Tmp21 and perinuclear targeting. Interestingly, isolated C1 domains from individual PKC isozymes differentially interact with p23/Tmp21. For PKCepsilon, it interacts with p23/Tmp21 specifically via its C1b domain; however, this association is lost in response to phorbol esters. These results demonstrate that p23/Tmp21 acts as an anchor that distinctively modulates compartmentalization of C1 domain-containing proteins, and it plays an essential role in beta2-chimaerin relocalization. Our study also highlights the relevance of C1 domains in protein-protein interactions in addition to their well-established lipid-binding properties.
Wang
Hongbin
H
Department of Pharmacology, University of Pennsylvania School of Medicine, Philadelphia, PA 19104-6160.
Kazanietz
Marcelo G
MG
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D020690:GTPase-Activating Proteins / Q000378:metabolism*; D018698:Glutamic Acid / Q000378:metabolism; D006801:Humans; D007930:Leucine / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000378:metabolism; D009363:Neoplasm Proteins / Q000737:chemistry* / Q000378:metabolism*; D029543:Nucleocytoplasmic Transport Proteins; D011485:Protein Binding; D011493:Protein Kinase C / Q000737:chemistry* / Q000378:metabolism*; D051744:Protein Kinase C-epsilon / Q000737:chemistry / Q000378:metabolism; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D034622:RNA Interference; D013329:Structure-Activity Relationship; D020744:rac GTP-Binding Proteins / Q000378:metabolism*; D000099221:Protein Kinase C zeta
D020690:GTPase-Activating Proteins; D018698:Glutamic Acid; D007930:Leucine; D008565:Membrane Proteins; D050505:Mutant Proteins; D009363:Neoplasm Proteins; D029543:Nucleocytoplasmic Transport Proteins; D011493:Protein Kinase C; D051744:Protein Kinase C-epsilon; D020744:rac GTP-Binding Proteins; D000099221:Protein Kinase C zeta; C484395:TMED10 protein, human; C079844:beta-chimaerin