JBC Anatrace, Inc.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Originally published In Press as doi:10.1074/jbc.M002203200 on May 5, 2000

J. Biol. Chem., Vol. 275, Issue 28, 21532-21538, July 14, 2000
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
275/28/21532    most recent
M002203200v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by van Tiel, C. M.
Right arrow Articles by Snoek, G. T.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by van Tiel, C. M.
Right arrow Articles by Snoek, G. T.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

The Protein Kinase C-dependent Phosphorylation of Serine 166 Is Controlled by the Phospholipid Species Bound to the Phosphatidylinositol Transfer Protein alpha *

Claudia M. van Tiel, Jan Westerman, Marten Paasman, Karel W. A. Wirtz, and Gerry T. SnoekDagger

From the Center for Biomembranes and Lipid Enzymology, Department of Lipid Biochemistry, Institute of Biomembranes, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands

The charge isomers of bovine brain PI-TPalpha (i.e. PI-TPalpha I containing a phosphatidylinositol (PI) molecule and PI-TPalpha II containing a phosphatidylcholine (PC) molecule) were phosphorylated in vitro by rat brain protein kinase C (PKC) at different rates. From the double-reciprocal plot, it was estimated that the Vmax values for PI-TPalpha I and II were 2.0 and 6.0 nmol/min, respectively; the Km values for both charge isomers were about equal, i.e. 0.7 µM. Phosphorylation of charge isomers of recombinant mouse PI-TPalpha confirmed that the PC-containing isomer was the better substrate. Phosphoamino acid analysis of in vitro and in vivo 32P-labeled PI-TPalpha s showed that serine was the major site of phosphorylation. Degradation of 32P-labeled PI-TPalpha by cyanogen bromide followed by high pressure liquid chromatography and sequence analysis yielded one 32P-labeled peptide (amino acids 104-190). This peptide contained Ser-148, Ser-152, and the consensus PKC phosphorylation site Ser-166. Replacement of Ser-166 with an alanine residue confirmed that indeed this residue was the site of phosphorylation. This mutation completely abolished PI and PC transfer activity. This was also observed when Ser-166 was replaced with Asp, implying that this is a key amino acid residue in regulating the function of PI-TPalpha . Stimulation of NIH3T3 fibroblasts by phorbol ester or platelet-derived growth factor induced the rapid relocalization of PI-TPalpha to perinuclear Golgi structures concomitant with a 2-3-fold increase in lysophosphatidylinositol levels. This relocalization was also observed for Myc-tagged wtPI-TPalpha expressed in NIH3T3 cells. In contrast, the distribution of Myc-tagged PI-TPalpha (S166A) and Myc-tagged PI-TPalpha (S166D) were not affected by phorbol ester, suggesting that phosphorylation of Ser-166 was a prerequisite for the relocalization to the Golgi. A model is proposed in which the PKC-dependent phosphorylation of PI-TPalpha is linked to the degradation of PI.

* This work was supported by the Netherlands Organization for Scientific Research.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Dagger To whom correspondence should be addressed. Tel.: 31-30-2534668; Fax: 31-30-2522478; E-mail: g.t.snoek@chem.uu.nl.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
Add to CiteULike CiteULike   Add to Complore Complore   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 Mol. Biol. CellHome page
D. Peretti, N. Dahan, E. Shimoni, K. Hirschberg, and S. Lev
Coordinated Lipid Transfer between the Endoplasmic Reticulum and the Golgi Complex Requires the VAP Proteins and Is Essential for Golgi-mediated Transport
Mol. Biol. Cell, September 1, 2008; 19(9): 3871 - 3884.
[Abstract] [Full Text] [PDF]

Home page
 Cardiovasc ResHome page
Y.-H. Yeh, J. R. Ehrlich, X. Qi, T. E. Hebert, D. Chartier, and S. Nattel
Adrenergic control of a constitutively active acetylcholine-regulated potassium current in canine atrial cardiomyocytes
Cardiovasc Res, June 1, 2007; 74(3): 406 - 415.
[Abstract] [Full Text] [PDF]

Home page
 Mol. Biol. CellHome page
S. E. Phillips, K. E. Ile, M. Boukhelifa, R. P.H. Huijbregts, and V. A. Bankaitis
Specific and Nonspecific Membrane-binding Determinants Cooperate in Targeting Phosphatidylinositol Transfer Protein beta-Isoform to the Mammalian Trans-Golgi Network
Mol. Biol. Cell, June 1, 2006; 17(6): 2498 - 2512.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
C. P. Morgan, A. Skippen, B. Segui, A. Ball, V. Allen-Baume, B. Larijani, J. Murray-Rust, N. McDonald, G. Sapkota, N. Morrice, et al.
Phosphorylation of a Distinct Structural Form of Phosphatidylinositol Transfer Protein {alpha} at Ser166 by Protein Kinase C Disrupts Receptor-mediated Phospholipase C Signaling by Inhibiting Delivery of Phosphatidylinositol to Membranes
J. Biol. Chem., November 5, 2004; 279(45): 47159 - 47171.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
L. Zhang, J.-K. Lee, S. A. John, N. Uozumi, and I. Kodama
Mechanosensitivity of GIRK Channels Is Mediated by Protein Kinase C-dependent Channel-Phosphatidylinositol 4,5-Bisphosphate Interaction
J. Biol. Chem., February 20, 2004; 279(8): 7037 - 7047.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
W.-Z. Zeng, X.-J. Li, D. W. Hilgemann, and C.-L. Huang
Protein Kinase C Inhibits ROMK1 Channel Activity via a Phosphatidylinositol 4,5-Bisphosphate-dependent Mechanism
J. Biol. Chem., May 2, 2003; 278(19): 16852 - 16856.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Cell Physiol.Home page
C. Nasuhoglu, S. Feng, Y. Mao, I. Shammat, M. Yamamato, S. Earnest, M. Lemmon, and D. W. Hilgemann
Modulation of cardiac PIP2 by cardioactive hormones and other physiologically relevant interventions
Am J Physiol Cell Physiol, July 1, 2002; 283(1): C223 - C234.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
C. M. van Tiel, J. Westerman, M. A. Paasman, M. M. Hoebens, K. W. A. Wirtz, and G. T. Snoek
The Golgi Localization of Phosphatidylinositol Transfer Protein beta Requires the Protein Kinase C-dependent Phosphorylation of Serine 262 and Is Essential for Maintaining Plasma Membrane Sphingomyelin Levels
J. Biol. Chem., June 14, 2002; 277(25): 22447 - 22452.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
M. D. Yoder, L. M. Thomas, J. M. Tremblay, R. L. Oliver, L. R. Yarbrough, and G. M. Helmkamp Jr.
Structure of a Multifunctional Protein. MAMMALIAN PHOSPHATIDYLINOSITOL TRANSFER PROTEIN COMPLEXED WITH PHOSPHATIDYLCHOLINE
J. Biol. Chem., March 16, 2001; 276(12): 9246 - 9252.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
 All ASBMB Journals   Molecular and Cellular Proteomics 
 Journal of Lipid Research   ASBMB Today 
Copyright © 2000 by the American Society for Biochemistry and Molecular Biology.