Protein Science Sheba protein
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Protein Science (2004), 13:1778-1786. Published by Cold Spring Harbor Laboratory Press. Copyright © 2004 The Protein Society
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
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 Deaton, J.
Right arrow Articles by Young, R.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Deaton, J.
Right arrow Articles by Young, R.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Solubilization and delivery by GroEL of megadalton complexes of the {lambda} holin

John Deaton1, Christos G. Savva2, Jingchuan Sun2, Andreas Holzenburg1,2, Joel Berry1 and Ry Young1

1 Department of Biochemistry and Biophysics and
2 Microscopy and Imaging Center and Department of Biology, Texas A&M University, College Station, Texas 77843, USA

(RECEIVED March 13, 2004; FINAL REVISION April 21, 2004; ACCEPTED April 21, 2004)



Abstract

GroEL can solubilize membrane proteins by binding them in its hydrophobic cavity when detergent is removed by dialysis. The best-studied example is bacteriorhodopsin, which can bind in the GroEL chaperonin at two molecules per tetradecamer. Applying this approach to the holin and antiholin proteins of phage {lambda}, we find that both proteins are solubilized by GroEL, in an ATP-sensitive mode, but to vastly different extents. The antiholin product, S107, saturates the chaperonin at six molecules per tetradecameric complex, whereas the holin, S105, which is missing the two N-terminal residues of S107, forms a hyper-solubilization complex with up to 350 holin molecules per GroEL, or approximately 4 MDa of protein per 0.8 MDa tetradecamer. Gel filtration chromatography and immunoprecipitation experiments confirmed the existence of complexes of the predicted masses for both S105 and S107 solubilization. For S105, negatively stained electron microscopic images show structures consistent with protein shells of the holin assembled around the chaperonin tetradecamer. Importantly, S105 can be delivered rapidly and efficiently to artificial liposomes from these complexes. In these delivery experiments, the holin exhibits efficient membrane-permeabilizing activity. The S107 antiholin can block formation of the hypersolubilization complexes, suggesting that their formation is related to an oligomerization step intrinsic to holin function.

Keywords: membrane proteins; chaperone; holins; liposomes

Abbreviations: BR, bacteriorhodopsin • BES, N,N'-bis(2-hydroxyethyl)-2-2aminoethanesulfonic acid • BSA, bovine serum albumin • DMSO, dimethylsulfoxide • EBB, Empigen BB • SDS-PAGE, sodium dodecyl sulfate polyacrylamide gel electrophoresis • TBS, Tris-buffered saline • TMD, transmembrane domain.

Reprint requests to: Ry Young, Department of Biochemistry and Biophysics, Texas A&M University, 2128 TAMU, College Station, TX 77843-2128, USA; e-mail: ryland{at}tamu.edu; fax: (979) 862-4718.

Article and publication are at http://www.proteinscience.org/cgi/doi/10.1110/ps.04735104.


Add to CiteULike CiteULike   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
 Proc. Natl. Acad. Sci. USAHome page
T. Park, D. K. Struck, J. F. Deaton, and R. Young
Topological dynamics of holins in programmed bacterial lysis
PNAS, December 26, 2006; 103(52): 19713 - 19718.
[Abstract] [Full Text] [PDF]

Home page
 J. Bacteriol.Home page
G. Ziedaite, R. Daugelavicius, J. K. H. Bamford, and D. H. Bamford
The Holin Protein of Bacteriophage PRD1 Forms a Pore for Small-Molecule and Endolysin Translocation
J. Bacteriol., August 1, 2005; 187(15): 5397 - 5405.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2004 by The Protein Society.