JCB logo
Avanti Polar Lipids, Inc.
  Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents
This Article
Right arrow Full Text
Right arrow Full Text (PDF, 824K)
Right arrow PPT slides of all figures
Right arrow Alert me when this article is cited
Right arrow Citation Map
Services
Right arrow Email this article
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new content in the JCB
Right arrow Download to citation manager
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via CrossRef
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Stan, R.-V.
Right arrow Articles by Palade, G. E.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Stan, R.-V.
Right arrow Articles by Palade, G. E.
Social Bookmarking
Add to CiteULike   Add to Complore   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Facebook   Add to Reddit   Add to Technorati   Add to Twitter  
What's this?

© The Rockefeller University Press, 0021-9525/1999//1189 $5.00
The Journal of Cell Biology, Volume 145, Number 6, , 1999 1189-1198

Article

Isolation, Cloning, and Localization of Rat PV-1, a Novel Endothelial Caveolar Protein



Radu-Virgil Stan*, Lucian Ghitescu{ddagger}, Bruce S. Jacobson§, and George E. Palade*

* Division of Cellular and Molecular Medicine, University of California, San Diego, San Diego, California 92093; {ddagger} Department of Pathology and Cell Biology, University of Montreal, Montreal, Canada H3C 1J4; and § Department of Biochemistry and Cell Biology, University of Massachusetts, Amherst, Massachusetts 01003

By using an immunoisolation procedure (Stan, R.-V., W.G. Roberts, K. Ihida, D. Predescu, L. Saucan, L. Ghitescu, and G.E. Palade. 1997. Mol. Biol. Cell. 8:595–605) developed in our laboratory, we have isolated a caveolar subfraction from rat lung endothelium and we have partially characterized the proteins of this subfraction which include an apparently caveolae-specific glycoprotein we propose to call PV-1 (formerly known as gp68). The isolation and partial sequencing of PV-1, combined with the cloning of the full length PV-1 cDNA led to the following conclusions: (a) PV-1 is a novel single span type II integral membrane protein (438 amino acids long) which forms homodimers in situ; (b) the transmembrane domain of PV-1 is near the NH2 terminus defining a short cytoplasmic endodomain and a large COOH-terminal ectodomain exposed to the blood plasma; (c) PV-1 is N-glycosylated and its glycan antennae bear terminal nonreducing galactosyl residues in {alpha}1-3 linkage. PV-1 is expressed mostly in the lung but both the messenger RNA and the protein can be detected at lower levels also in kidney, spleen, liver, heart, muscle, and brain. No signal could be detected in testis and two lower molecular weight forms were detected in brain. Immunocytochemical studies carried out by immunodiffusion on rat lung with an anti–PV-1 polyclonal antibody directed against a COOH-terminal epitope reveal a specific localization of PV-1 to the stomatal diaphragms of rat lung endothelial caveolae and confirm the extracellular orientation of the PV-1 COOH terminus.

Key Words: plasmalemmal vesicles • Griffonia simplicifolia • lectin chromatography • rat lung • immunodiffusion

Abbreviations used in this paper: aa, amino acid; GS I, Griffonia simplicifolia I lectin; PBSA, 1% BSA in PBS; PEG, polyethylene glycol; PLP, paraformaldehyde-lysine-metaperiodate fixate; PV, plasmalemmal vesicles; PVDF, polyvinyldifluoride; RT, room temperature.

Part of this work was submitted in abstract form to the 38th American Society for Cell Biology annual meeting, December 12–16, San Francisco (Mol. Biol. Cell. 9[Suppl.].96a).


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 Facebook Facebook   Add to Reddit Reddit   Add to Technorati Technorati   Add to Twitter Twitter    What's this?


This article has been cited by other articles:



  Home | Help | Feedback | Subscriptions | Archive | Search | Table of Contents