HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

J. Biol. Chem., Vol. 279, Issue 7, 5207-5215, February 13, 2004

This Article Full Text Full Text (PDF) All Versions of this Article: 279/7/5207    most recent M308819200v1 Submit a Letter to Editor Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Rünzler, D. Articles by Sára, M. Search for Related Content PubMed PubMed Citation Articles by Rünzler, D. Articles by Sára, M. Social Bookmarking                      What's this?

Biophysical Characterization of the Entire Bacterial Surface Layer Protein SbsB and Its Two Distinct Functional Domains^*

Dominik Rünzler¶, Carina Huber||**, Dieter Moll||, Gottfried Köhler¶, and Margit Sára||

From the Institute for Theoretical Chemistry and Structural Biology, University of Vienna, Campus Vienna Biocenter 6/1, Vienna 1030, the ^¶Austrian Society for Aerospace Medicine, Lustkandlgasse 52/3, Vienna 1090, the ^||Center for Ultrastructure Research and Ludwig Boltzmann Institute for Molecular Nanotechnology, BOKU, University of Natural Resources and Applied Life Sciences, Gregor Mendelstrasse 33, Vienna 1180, and ^**Biomolecular Therapeutics GmbH, Brunnerstrasse 59, Vienna 1235, Austria

The crystalline bacterial cell surface layer (S-layer) protein SbsB of Geobacillus stearothermophilus PV72/p2 was dissected into an N-terminal part defined by the three consecutive S-layer homologous motifs and the remaining large C-terminal part. Both parts of the mature protein were produced as separate recombinant proteins (rSbsB_1–178 and rSbsB_177–889) and compared with the full-length form rSbsB_1–889 (rSbsB). Evidence for functional and structural integrity of the two truncated forms was provided by optical spectroscopic methods and electron microscopy. In particular, binding of the secondary cell wall polymer revealed a high affinity dissociation constant of 3 nM and could be assigned solely to the soluble rSbsB_1–178, whereas rSbsB_177–889 self-assembled into the same lattice as the full-length protein. Furthermore, thermal as well as guanidinium hydrochloride induced equilibrium unfolding profiles monitored by intrinsic fluorescence, and circular dichroism spectroscopy allowed characterization of rSbsB_1–178 as an -helical protein with a single cooperative unfolding transition yielding a G value of 26.5 kJ mol^–1. The C-terminal rSbsB_177–889 could be characterized as a -sheet protein with typical multidomain unfolding, which is partially less stable as stand-alone protein. In general, the truncated forms showed identical properties compared with the full-length rSbsB with respect to structure and function. Consequently, rSbsB is characterized by its two functionally and structurally separated parts, the specific secondary cell wall polymer binding rSbsB_1–178 and the larger rSbsB_177–889 responsible for formation of the crystalline array.

Received for publication, August 11, 2003 , and in revised form, November 17, 2003.

^* This work was supported by the Austrian Science Foundation, Project 14689, and the Competence Center "Biomolecular Therapeutics." The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

Both authors contributed equally to this work.

To whom correspondence should be addressed: Institute for Theoretical Chemistry and Structural Biology, University of Vienna, Campus Vienna Biocenter 6/1, Vienna 1030, Austria. Tel.: 43-1-4277-53012; Fax: 43-1-4277-53099; E-mail: dominik.ruenzler{at}univie.ac.at.

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				J. Ferner-Ortner, C. Mader, N. Ilk, U. B. Sleytr, and E. M. Egelseer High-Affinity Interaction between the S-Layer Protein SbsC and the Secondary Cell Wall Polymer of Geobacillus stearothermophilus ATCC 12980 Determined by Surface Plasmon Resonance Technology J. Bacteriol., October 1, 2007; 189(19): 7154 - 7158. [Abstract] [Full Text] [PDF]

				T. Candela, T. Mignot, X. Hagnerelle, M. Haustant, and A. Fouet Genetic analysis of Bacillus anthracis Sap S-layer protein crystallization domain Microbiology, May 1, 2005; 151(5): 1485 - 1490. [Abstract] [Full Text] [PDF]