TRP1 interacting PDZ-domain protein GIPC forms oligomers and is localized to intracellular vesicles in human melanocytes

Rajendra H. Kedlaya; Kumar M.R. Bhat; Julie Mitchell; Steven J. Darnell; Vijayasaradhi Setaluri

doi:10.1016/j.abb.2006.08.010

TRP1 interacting PDZ-domain protein GIPC forms oligomers and is localized to intracellular vesicles in human melanocytes

Rajendra H. Kedlaya, Kumar M.R. Bhat, Julie Mitchell, Steven J. Darnell, Vijayasaradhi Setaluri

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

11 Scopus citations

Abstract

PDZ proteins coordinate assembly of protein complexes that participate in diverse biological processes. GIPC is a multifunctional PDZ protein that interacts with several soluble and membrane proteins. Unlike most PDZ proteins, GIPC contains single PDZ domain and the mechanisms by which GIPC mediates its actions remain unclear. We investigated the possibility that in lieu of multiple PDZ domains, GIPC forms multimers. Here, we demonstrate that GIPC can bind to itself and that the PDZ domain is involved in GIPC-GIPC interaction. Gel filtration, sucrose gradient centrifugation and chemical cross-linking showed that whereas bulk of cytosolic GIPC was present as monomer, oligomers with an estimated molecular mass corresponding to GIPC homotrimer were readily detectable in the membrane fraction. Modeling of GIPC PDZ domain showed feasibility of trimerization. Immunogold electron microscopy showed that GIPC is present in clusters near vesicles. Our data suggest that oligomers of GIPC mediate its functions in melanocytes.

Original language	English
Pages (from-to)	160-169
Number of pages	10
Journal	Archives of Biochemistry and Biophysics
Volume	454
Issue number	2
DOIs	https://doi.org/10.1016/j.abb.2006.08.010
State	Published - Oct 15 2006
Externally published	Yes

Funding

We thank Dr. Michael F. Hoffman, University of Wisconsin, for providing GST-thioredoxin fusion protein construct and Dr. Raymond Boissy, University of Cincinnati, for help with electron microscopy. This work was supported by NIH Grant R01AR048913.

Funders	Funder number
National Institutes of Health
National Institute of Arthritis and Musculoskeletal and Skin Diseases	R01AR048913

Keywords

Chemical cross-linking
Computational analysis
GIPC
Oligomerization
PDZ proteins
Protein folding

Access to Document

10.1016/j.abb.2006.08.010

Cite this

@article{b88c2c165b5d4a7abcd4597ab6fe167d,

title = "TRP1 interacting PDZ-domain protein GIPC forms oligomers and is localized to intracellular vesicles in human melanocytes",

abstract = "PDZ proteins coordinate assembly of protein complexes that participate in diverse biological processes. GIPC is a multifunctional PDZ protein that interacts with several soluble and membrane proteins. Unlike most PDZ proteins, GIPC contains single PDZ domain and the mechanisms by which GIPC mediates its actions remain unclear. We investigated the possibility that in lieu of multiple PDZ domains, GIPC forms multimers. Here, we demonstrate that GIPC can bind to itself and that the PDZ domain is involved in GIPC-GIPC interaction. Gel filtration, sucrose gradient centrifugation and chemical cross-linking showed that whereas bulk of cytosolic GIPC was present as monomer, oligomers with an estimated molecular mass corresponding to GIPC homotrimer were readily detectable in the membrane fraction. Modeling of GIPC PDZ domain showed feasibility of trimerization. Immunogold electron microscopy showed that GIPC is present in clusters near vesicles. Our data suggest that oligomers of GIPC mediate its functions in melanocytes.",

keywords = "Chemical cross-linking, Computational analysis, GIPC, Oligomerization, PDZ proteins, Protein folding",

author = "Kedlaya, {Rajendra H.} and Bhat, {Kumar M.R.} and Julie Mitchell and Darnell, {Steven J.} and Vijayasaradhi Setaluri",

year = "2006",

month = oct,

day = "15",

doi = "10.1016/j.abb.2006.08.010",

language = "English",

volume = "454",

pages = "160--169",

journal = "Archives of Biochemistry and Biophysics",

issn = "0003-9861",

publisher = "Elsevier",

number = "2",

}

TY - JOUR

T1 - TRP1 interacting PDZ-domain protein GIPC forms oligomers and is localized to intracellular vesicles in human melanocytes

AU - Kedlaya, Rajendra H.

AU - Bhat, Kumar M.R.

AU - Mitchell, Julie

AU - Darnell, Steven J.

AU - Setaluri, Vijayasaradhi

PY - 2006/10/15

Y1 - 2006/10/15

N2 - PDZ proteins coordinate assembly of protein complexes that participate in diverse biological processes. GIPC is a multifunctional PDZ protein that interacts with several soluble and membrane proteins. Unlike most PDZ proteins, GIPC contains single PDZ domain and the mechanisms by which GIPC mediates its actions remain unclear. We investigated the possibility that in lieu of multiple PDZ domains, GIPC forms multimers. Here, we demonstrate that GIPC can bind to itself and that the PDZ domain is involved in GIPC-GIPC interaction. Gel filtration, sucrose gradient centrifugation and chemical cross-linking showed that whereas bulk of cytosolic GIPC was present as monomer, oligomers with an estimated molecular mass corresponding to GIPC homotrimer were readily detectable in the membrane fraction. Modeling of GIPC PDZ domain showed feasibility of trimerization. Immunogold electron microscopy showed that GIPC is present in clusters near vesicles. Our data suggest that oligomers of GIPC mediate its functions in melanocytes.

AB - PDZ proteins coordinate assembly of protein complexes that participate in diverse biological processes. GIPC is a multifunctional PDZ protein that interacts with several soluble and membrane proteins. Unlike most PDZ proteins, GIPC contains single PDZ domain and the mechanisms by which GIPC mediates its actions remain unclear. We investigated the possibility that in lieu of multiple PDZ domains, GIPC forms multimers. Here, we demonstrate that GIPC can bind to itself and that the PDZ domain is involved in GIPC-GIPC interaction. Gel filtration, sucrose gradient centrifugation and chemical cross-linking showed that whereas bulk of cytosolic GIPC was present as monomer, oligomers with an estimated molecular mass corresponding to GIPC homotrimer were readily detectable in the membrane fraction. Modeling of GIPC PDZ domain showed feasibility of trimerization. Immunogold electron microscopy showed that GIPC is present in clusters near vesicles. Our data suggest that oligomers of GIPC mediate its functions in melanocytes.

KW - Chemical cross-linking

KW - Computational analysis

KW - GIPC

KW - Oligomerization

KW - PDZ proteins

KW - Protein folding

UR - http://www.scopus.com/inward/record.url?scp=33749619076&partnerID=8YFLogxK

U2 - 10.1016/j.abb.2006.08.010

DO - 10.1016/j.abb.2006.08.010

M3 - Article

C2 - 16962991

AN - SCOPUS:33749619076

SN - 0003-9861

VL - 454

SP - 160

EP - 169

JO - Archives of Biochemistry and Biophysics

JF - Archives of Biochemistry and Biophysics

IS - 2

ER -

TRP1 interacting PDZ-domain protein GIPC forms oligomers and is localized to intracellular vesicles in human melanocytes

Abstract

Funding

Keywords

Access to Document

Other files and links

Fingerprint

Cite this