![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
![[Contents]](/icons/banner/tocACT.gif){kind=icon}
|
|
|
|
|
||
Vol. 17, Issue 1, 56-66, January 2006
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
to the Plasma Membrane Requires Both Ca2+ and PIP2 Recognition by Its C2 Domain
* Molecular Biophysics Program and Department of Chemistry and Biochemistry, University of Colorado, Boulder, Boulder, CO 80309-0215;
Department of Microbiology and Immunology, Weill Medical College of Cornell University, New York, NY 10021;
Department of Pediatrics, National Jewish Medical and Research Center, Denver, CO 80206; and
Departments of Pathology and Pharmacology, University of Colorado School of Medicine, Denver, CO 80262
Submitted June 7, 2005;
Revised September 20, 2005;
Accepted October 12, 2005
Monitoring Editor: John York
The C2 domain of protein kinase C
(PKC) controls the translocation of this kinase from the cytoplasm to the plasma membrane during cytoplasmic Ca2+ signals. The present study uses intracellular coimaging of fluorescent fusion proteins and an in vitro FRET membrane-binding assay to further investigate the nature of this translocation. We find that Ca2+-activated PKC and its isolated C2 domain localize exclusively to the plasma membrane in vivo and that a plasma membrane lipid, phosphatidylinositol-4,5-bisphosphate (PIP2), dramatically enhances the Ca2+-triggered binding of the C2 domain to membranes in vitro. Similarly, a hybrid construct substituting the PKC Ca2+-binding loops (CBLs) and PIP2 binding site (
-strands 3–4) into a different C2 domain exhibits native Ca2+-triggered targeting to plasma membrane and recognizes PIP2. Conversely, a hybrid containing the CBLs but lacking the PIP2 site translocates primarily to trans-Golgi network (TGN) and fails to recognize PIP2. Similarly, PKC C2 domains possessing mutations in the PIP2 site target primarily to TGN and fail to recognize PIP2. Overall, these findings demonstrate that the CBLs are essential for Ca2+-triggered membrane binding but are not sufficient for specific plasma membrane targeting. Instead, targeting specificity is provided by basic residues on -strands 3–4, which bind to plasma membrane PIP2.
The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).
Address correspondence to: John H. Evans (John.Evans{at}colorado.edu) or Joseph J. Falke (Joseph.Falke{at}colorado.edu).
This article has been cited by other articles:
|
|
 |
|
  M. Guerrero-Valero, C. Ferrer-Orta, J. Querol-Audi, C. Marin-Vicente, I. Fita, J. C. Gomez-Fernandez, N. Verdaguer, and S. Corbalan-Garcia Structural and mechanistic insights into the association of PKC{alpha}-C2 domain to PtdIns(4,5)P2 PNAS, April 21, 2009; 106(16): 6603 - 6607. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. B. Lee, S. Kim, and B. McClure A Pollen Protein, NaPCCP, That Binds Pistil Arabinogalactan Proteins Also Binds Phosphatidylinositol 3-Phosphate and Associates with the Pollen Tube Endomembrane System Plant Physiology, February 1, 2009; 149(2): 791 - 802. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 N. Coudevylle, P. Montaville, A. Leonov, M. Zweckstetter, and S. Becker Structural Determinants for Ca2+ and Phosphatidylinositol 4,5-Bisphosphate Binding by the C2A Domain of Rabphilin-3A J. Biol. Chem., December 19, 2008; 283(51): 35918 - 35928. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. F. Steinberg Structural Basis of Protein Kinase C Isoform Function Physiol Rev, October 1, 2008; 88(4): 1341 - 1378. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
D. Manna, N. Bhardwaj, M. S. Vora, R. V. Stahelin, H. Lu, and W. Cho Differential Roles of Phosphatidylserine, PtdIns(4,5)P2, and PtdIns(3,4,5)P3 in Plasma Membrane Targeting of C2 Domains: MOLECULAR DYNAMICS SIMULATION, MEMBRANE BINDING, AND CELL TRANSLOCATION STUDIES OF THE PKC{alpha} C2 Domain J. Biol. Chem., September 19, 2008; 283(38): 26047 - 26058. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. H. Evans and J. J. Falke Ca2+ influx is an essential component of the positive-feedback loop that maintains leading-edge structure and activity in macrophages PNAS, October 9, 2007; 104(41): 16176 - 16181. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S.-W. Kim, J. S. Hong, S.-H. Ryu, W.-C. Chung, J.-H. Yoon, and J. S. Koo Regulation of Mucin Gene Expression by CREB via a Nonclassical Retinoic Acid Signaling Pathway Mol. Cell. Biol., October 1, 2007; 27(19): 6933 - 6947. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
H. Stensman and C. Larsson Identification of Acidic Amino Acid Residues in the Protein Kinase C{alpha} V5 Domain That Contribute to Its Insensitivity to Diacylglycerol J. Biol. Chem., September 28, 2007; 282(39): 28627 - 28638. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
R. V. Stahelin, P. Subramanian, M. Vora, W. Cho, and C. E. Chalfant Ceramide-1-phosphate Binds Group IVA Cytosolic Phospholipase a2 via a Novel Site in the C2 Domain J. Biol. Chem., July 13, 2007; 282(28): 20467 - 20474. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. L. Veale, L. E. Kennard, G. L. Sutton, G. MacKenzie, C. Sandu, and A. Mathie G{alpha}q-Mediated Regulation of TASK3 Two-Pore Domain Potassium Channels: The Role of Protein Kinase C Mol. Pharmacol., June 1, 2007; 71(6): 1666 - 1675. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
O. Rey, S. H. Young, R. Papazyan, M. S. Shapiro, and E. Rozengurt Requirement of the TRPC1 Cation Channel in the Generation of Transient Ca2+ Oscillations by the Calcium-sensing Receptor J. Biol. Chem., December 15, 2006; 281(50): 38730 - 38737. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. E. Dowd, S. Coursol, A. L. Skirpan, T.-h. Kao, and S. Gilroy Petunia Phospholipase C1 Is Involved in Pollen Tube Growth PLANT CELL, June 1, 2006; 18(6): 1438 - 1453. [Abstract] [Full Text] [PDF]
|
|
