Abstract
The BspA protein of Lactobacillus fermentum BR11 (BR11) is a cell envelope constituent that is similar to known solute-binding proteins and putative adhesins. BspA is required for L-cystine uptake and oxidative defense and is likely to be an L-cystine-binding protein. The aim of this study was to directly measure L-cystine-BspA binding and BspA expression. De-energized BR11 cells bound radiolabelled L-cystine with a Kd of 0.2 μM. A bspA mutant could not bind L-cystine. L-cystine-BR11 binding was unaffected by large excesses of L-glutamine, L-methionine, or collagen, indicating L-cystine specificity. BR11 and the bspA mutant were identical in their abilities to bind L-cysteine, indicating that L-cysteine is not a BspA ligand. BspA expression levels were deduced from radiolabelled L-cystine binding and it was found that there are 1–2 × 105 BspA molecules per cell, and that expression is slightly higher under oxidizing conditions. It is proposed that BspA be renamed CyuC.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Literature Cited
BS Gan J Kim G Reid P Cadieux JC Howard (2002) ArticleTitleLactobacillus fermentum RC-14 inhibits Staphylococcus aureus infection of surgical implants in rats J Infect Dis. 185 1369–1372
CEG Havenith JFML Seegers PH Pouwels (2002) ArticleTitleGut associated lactobacilli for oral immunisation Food Res Int. 35 151–163
C Heinemann JE Hylckama Vlieg Particlevan DB Janssen HJ Busscher HC Mei Particlevan der G Reid (2000) ArticleTitlePurification and characterization of a surface-binding protein from Lactobacillus fermentum RC-14 that inhibits adhesion of Enterococcus faecalis 1131 FEMS Microbiol Lett. 190 177–180
J Hung D Cooper MS Turner T Walsh PM Giffard (2003) ArticleTitleCystine uptake prevents production of hydrogen peroxide by Lactobacillus fermentum BR11 FEMS Microbiol Lett. 227 93–99
HF Jenkinson RA Baker GW Tannock (1996) ArticleTitleA binding-lipoprotein-dependent oligopeptide transport system in Streptococcus gordonii essential for uptake of hexa- and heptapeptides J Bacteriol. 178 68–77
UK Laemmli M Favre (1973) ArticleTitleMaturation of the head of bacteriophage T4. I DNA packaging events J Mol Biol. 80 575–599
DM Miller Suffix3rd JS Olson JW Pflugrath FA Ouiocho (1983) ArticleTitleRates of ligand binding to periplasmic proteins involved in bacterial transport and chemotaxis J Biol Chem. 258 13665–13672
J Mills NR Wyborn JA Greenwood SG Williams CW Jones (1998) ArticleTitleCharacterisation of a binding-protein-dependent, active transport system for short-chain amides and urea in the methylotrophic bacterium Methylophilus methylotrophus Eur J Biochem. 251 45–53
M Pajatsch M Gerhart R Peist R Horlacher W Boos A Bock (1998) ArticleTitleThe periplasmic cyclodextrin binding protein CymE from Klebsiella oxytoca and its role in maltodextrin and cyclodextrin transport J Bacteriol. 180 2630–2635
Z Pei C Burucoa B Grignon S Bagar XZ Huang DJ Kopecko AL Bourgeois JL Fouchere MJ Bloser (1998) ArticleTitleMutation in the peb1A locus of Campylobacter jejuni reduces interactions with epithelial cells and intestinal colonization of mice Infect Immun. 66 938–943
M Rojas F Ascencio PL Conway (2002) ArticleTitlePurification and characterization of a surface protein from Lactobacillus fermentum 104R that binds to porcine small intestinal mucus and gastric mucin Appl Environ Microbiol. 68 2330–2336
S Roos P Aleljung N Robert B Lee T Wadstrom M Lindberg H Jonsson (1996) ArticleTitleA collagen binding protein from Lactobacillus reuteri is part of an ABC transporter system? FEMS Microbiol Lett. 144 33–38
CM Rush LM Hafner P Timms (1994) ArticleTitleGenetic modification of a vaginal strain of Lactobacillus fermentum and its maintenance within the reproductive tract after intravaginal administration J Med Microbiol. 41 272–278
J Sagara K Miura S Bannai (1993) ArticleTitleCystine uptake and glutathione level in fetal brain cells in primary culture and in suspension J Neurochem. 61 1667–1671
UB Sleytr P Messner (1988) ArticleTitleCrystalline surface layers in procaryotes J Bacteriol. 170 2891–2897
I Susanto SE Wright RS Lawson CE Williams SM Deneke (1998) ArticleTitleMetallothionein, glutathione, and cystine transport in pulmonary artery endothelial cells and NIH/3T3 cells Am J Physiol. 274 L296–L300
R Tam MH Saier SuffixJr (1993) ArticleTitleStructural, functional, and evolutionary relationships among extracellular solute-binding receptors of bacteria Microbiol Rev. 57 320–346
MS Turner P Timms LM Hafner PM Giffard (1997) ArticleTitleIdentification and characterization of a basic cell surface-located protein from Lactobacillus fermentum BR11 J Bacteriol. 179 3310–3316
MS Turner T Woodberry LM Hafner PM Giffard (1999) ArticleTitleThe bspA locus of Lactobacillus fermentum BR11 encodes an L-cystine uptake system J Bacteriol. 181 2192–2198
Acknowledgments
This work was supported in part by a QUT special research grant. J.H. was supported by a QUT postgraduate scholarship.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Hung, J., Turner, M.S., Walsh, T. et al. BspA (CyuC) in Lactobacillus fermentum BR11 Is a Highly Expressed High-Affinity L-Cystine-Binding Protein. Curr Microbiol 50, 33–37 (2005). https://doi.org/10.1007/s00284-004-4408-2
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00284-004-4408-2