Structural basis of syndecan-4 phosphorylation as a molecular switch to regulate signaling

Bon Kyung Koo; Young Sang Jung; Joon Shin; Innoc Han; Eva Mortier; Pascale Zimmermann; James R. Whiteford; John R. Couchman; Eok Soo Oh; Weontae Lee

doi:10.1016/j.jmb.2005.09.087

Structural basis of syndecan-4 phosphorylation as a molecular switch to regulate signaling

Bon Kyung Koo, Young Sang Jung, Joon Shin, Innoc Han, Eva Mortier, Pascale Zimmermann, James R. Whiteford, John R. Couchman, Eok Soo Oh, Weontae Lee

Department of Life Science

Research output: Contribution to journal › Article › peer-review

64 Scopus citations

Abstract

The syndecan transmembrane proteoglycans are involved in the organization of the actin cytoskeleton and have important roles as cell surface receptors during cell-matrix interactions. We have shown that the syndecan-4 cytoplasmic domain (4L) forms oligomeric complexes that bind to and stimulate PKCα activity in the presence of PtdIns(4,5)P₂, emphasizing the importance of multimerization in the regulation of PKCα activation. Oligomerization of the cytoplasmic domain of syndecan-4 is regulated either positively by PtdIns(4,5)P₂ or negatively by phosphorylation of serine 183. Phosphorylation results in reduced PKCα activity by inhibiting PtdIns(4,5)P₂-dependent oligomerization of the syndecan-4 cytoplasmic domain. Data from NMR and gel-filtration chromatography show that the phosphorylated cytoplasmic domain (p-4L) exists as a dimer, similar to 4L, but not as higher-order oligomers. NMR analysis showed that the overall conformation of p-4L is a compact intertwined dimer with an unusually symmetric clamp shape, and its molecular surface is mostly positively charged. The two parallel strands form a cavity in the center of the dimeric twist. An especially marked effect of phosphorylation of the syndecan-4 cytoplasmic domain is a dramatic conformational change near the C2 region that ablates an interaction site with the PDZ domain of syntenin. Wound healing studies further suggest that syndecan-4 phosphorylation might influence cell migration behavior. We conclude that the phosphorylation (Ser183) of syndecan-4 can play a critical role as a molecular switch to regulate its functions through conformational change.

Original language	English
Pages (from-to)	651-663
Number of pages	13
Journal	Journal of Molecular Biology
Volume	355
Issue number	4
DOIs	https://doi.org/10.1016/j.jmb.2005.09.087
State	Published - 27 Jan 2006

Bibliographical note

Funding Information:
This work was supported by the NRL program of MOST NRDP (M1-0203-00-0020) (to W.L.) Korean Research Foundation grants (KRF-99-042-D00096, KRF-2002-CP0327) and the Korea Science and Engineering Foundation (KOSEF) through the Protein Network Research Center at Yonsei University (to W.L.). This work is supported, in part, by the Brain Korea 21 Project. J.R.C. and J.R.W. are supported by Wellcome Trust Programme Grant 065940. P.Z. and E.M. were supported by The Flanders Interuniversity Institute for Biotechnology, the Concerted Actions Program (GOA/2001/06) and by a grant from the Fund for Scientific Research, Flanders. P.Z. is a Postdoctoral Fellow of the Fund for Scientific Research, Flanders, Belgium (F.W.O.).

Keywords

NMR
Phosphatidylinositol 4, 5-bisphosphate
Phosphorylation
Solution structure
Syndecan-4

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title = "Structural basis of syndecan-4 phosphorylation as a molecular switch to regulate signaling",

abstract = "The syndecan transmembrane proteoglycans are involved in the organization of the actin cytoskeleton and have important roles as cell surface receptors during cell-matrix interactions. We have shown that the syndecan-4 cytoplasmic domain (4L) forms oligomeric complexes that bind to and stimulate PKCα activity in the presence of PtdIns(4,5)P2, emphasizing the importance of multimerization in the regulation of PKCα activation. Oligomerization of the cytoplasmic domain of syndecan-4 is regulated either positively by PtdIns(4,5)P2 or negatively by phosphorylation of serine 183. Phosphorylation results in reduced PKCα activity by inhibiting PtdIns(4,5)P2-dependent oligomerization of the syndecan-4 cytoplasmic domain. Data from NMR and gel-filtration chromatography show that the phosphorylated cytoplasmic domain (p-4L) exists as a dimer, similar to 4L, but not as higher-order oligomers. NMR analysis showed that the overall conformation of p-4L is a compact intertwined dimer with an unusually symmetric clamp shape, and its molecular surface is mostly positively charged. The two parallel strands form a cavity in the center of the dimeric twist. An especially marked effect of phosphorylation of the syndecan-4 cytoplasmic domain is a dramatic conformational change near the C2 region that ablates an interaction site with the PDZ domain of syntenin. Wound healing studies further suggest that syndecan-4 phosphorylation might influence cell migration behavior. We conclude that the phosphorylation (Ser183) of syndecan-4 can play a critical role as a molecular switch to regulate its functions through conformational change.",

keywords = "NMR, Phosphatidylinositol 4, 5-bisphosphate, Phosphorylation, Solution structure, Syndecan-4",

author = "Koo, {Bon Kyung} and Jung, {Young Sang} and Joon Shin and Innoc Han and Eva Mortier and Pascale Zimmermann and Whiteford, {James R.} and Couchman, {John R.} and Oh, {Eok Soo} and Weontae Lee",

note = "Funding Information: This work was supported by the NRL program of MOST NRDP (M1-0203-00-0020) (to W.L.) Korean Research Foundation grants (KRF-99-042-D00096, KRF-2002-CP0327) and the Korea Science and Engineering Foundation (KOSEF) through the Protein Network Research Center at Yonsei University (to W.L.). This work is supported, in part, by the Brain Korea 21 Project. J.R.C. and J.R.W. are supported by Wellcome Trust Programme Grant 065940. P.Z. and E.M. were supported by The Flanders Interuniversity Institute for Biotechnology, the Concerted Actions Program (GOA/2001/06) and by a grant from the Fund for Scientific Research, Flanders. P.Z. is a Postdoctoral Fellow of the Fund for Scientific Research, Flanders, Belgium (F.W.O.). ",

year = "2006",

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doi = "10.1016/j.jmb.2005.09.087",

language = "English",

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journal = "Journal of Molecular Biology",

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T1 - Structural basis of syndecan-4 phosphorylation as a molecular switch to regulate signaling

AU - Koo, Bon Kyung

AU - Jung, Young Sang

AU - Shin, Joon

AU - Han, Innoc

AU - Mortier, Eva

AU - Zimmermann, Pascale

AU - Whiteford, James R.

AU - Couchman, John R.

AU - Oh, Eok Soo

AU - Lee, Weontae

N1 - Funding Information: This work was supported by the NRL program of MOST NRDP (M1-0203-00-0020) (to W.L.) Korean Research Foundation grants (KRF-99-042-D00096, KRF-2002-CP0327) and the Korea Science and Engineering Foundation (KOSEF) through the Protein Network Research Center at Yonsei University (to W.L.). This work is supported, in part, by the Brain Korea 21 Project. J.R.C. and J.R.W. are supported by Wellcome Trust Programme Grant 065940. P.Z. and E.M. were supported by The Flanders Interuniversity Institute for Biotechnology, the Concerted Actions Program (GOA/2001/06) and by a grant from the Fund for Scientific Research, Flanders. P.Z. is a Postdoctoral Fellow of the Fund for Scientific Research, Flanders, Belgium (F.W.O.).

PY - 2006/1/27

Y1 - 2006/1/27

N2 - The syndecan transmembrane proteoglycans are involved in the organization of the actin cytoskeleton and have important roles as cell surface receptors during cell-matrix interactions. We have shown that the syndecan-4 cytoplasmic domain (4L) forms oligomeric complexes that bind to and stimulate PKCα activity in the presence of PtdIns(4,5)P2, emphasizing the importance of multimerization in the regulation of PKCα activation. Oligomerization of the cytoplasmic domain of syndecan-4 is regulated either positively by PtdIns(4,5)P2 or negatively by phosphorylation of serine 183. Phosphorylation results in reduced PKCα activity by inhibiting PtdIns(4,5)P2-dependent oligomerization of the syndecan-4 cytoplasmic domain. Data from NMR and gel-filtration chromatography show that the phosphorylated cytoplasmic domain (p-4L) exists as a dimer, similar to 4L, but not as higher-order oligomers. NMR analysis showed that the overall conformation of p-4L is a compact intertwined dimer with an unusually symmetric clamp shape, and its molecular surface is mostly positively charged. The two parallel strands form a cavity in the center of the dimeric twist. An especially marked effect of phosphorylation of the syndecan-4 cytoplasmic domain is a dramatic conformational change near the C2 region that ablates an interaction site with the PDZ domain of syntenin. Wound healing studies further suggest that syndecan-4 phosphorylation might influence cell migration behavior. We conclude that the phosphorylation (Ser183) of syndecan-4 can play a critical role as a molecular switch to regulate its functions through conformational change.

AB - The syndecan transmembrane proteoglycans are involved in the organization of the actin cytoskeleton and have important roles as cell surface receptors during cell-matrix interactions. We have shown that the syndecan-4 cytoplasmic domain (4L) forms oligomeric complexes that bind to and stimulate PKCα activity in the presence of PtdIns(4,5)P2, emphasizing the importance of multimerization in the regulation of PKCα activation. Oligomerization of the cytoplasmic domain of syndecan-4 is regulated either positively by PtdIns(4,5)P2 or negatively by phosphorylation of serine 183. Phosphorylation results in reduced PKCα activity by inhibiting PtdIns(4,5)P2-dependent oligomerization of the syndecan-4 cytoplasmic domain. Data from NMR and gel-filtration chromatography show that the phosphorylated cytoplasmic domain (p-4L) exists as a dimer, similar to 4L, but not as higher-order oligomers. NMR analysis showed that the overall conformation of p-4L is a compact intertwined dimer with an unusually symmetric clamp shape, and its molecular surface is mostly positively charged. The two parallel strands form a cavity in the center of the dimeric twist. An especially marked effect of phosphorylation of the syndecan-4 cytoplasmic domain is a dramatic conformational change near the C2 region that ablates an interaction site with the PDZ domain of syntenin. Wound healing studies further suggest that syndecan-4 phosphorylation might influence cell migration behavior. We conclude that the phosphorylation (Ser183) of syndecan-4 can play a critical role as a molecular switch to regulate its functions through conformational change.

KW - NMR

KW - Phosphatidylinositol 4, 5-bisphosphate

KW - Phosphorylation

KW - Solution structure

KW - Syndecan-4

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DO - 10.1016/j.jmb.2005.09.087

M3 - Article

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JO - Journal of Molecular Biology

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IS - 4

ER -

Structural basis of syndecan-4 phosphorylation as a molecular switch to regulate signaling

Abstract

Bibliographical note

Keywords

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