title: Surface (S)-layer proteins of Deinococcus radiodurans and their utility as vehicles for surface localization of functional proteins.
abstract: The radiation resistant bacterium, Deinococcus radiodurans contains two major surface (S)-layer proteins, Hpi and SlpA. The Hpi protein was shown to (a) undergo specific in vivo cleavage, and (b) closely associate with the SlpA protein. Using a non-specific acid phosphatase from Salmonella enterica serovar Typhi, PhoN as a reporter, the Surface Layer Homology (SLH) domain of SlpA was shown to bind deinococcal peptidoglycan-containing cell wall sacculi. The association of SlpA with Hpi on one side and peptidoglycan on the other, localizes this protein in the 'interstitial' layer of the deinoccocal cell wall. Gene chimeras of hpi-phoN and slh-phoN were constructed to test efficacy of S-layer proteins, as vehicles for cell surface localization in D. radiodurans. The Hpi-PhoN protein localized exclusively in the membrane fraction, and displayed cell-based phosphatase activity in vivo. The SLH-PhoN, which localized to both cytosolic and membrane fractions, displayed in vitro activity but no cell-based in vivo activity. Hpi, therefore, emerged as an efficient surface localizing protein and can be exploited for suitable applications of this superbug.
authors.affiliation: Molecular Biology Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085, India.
authors.forename: Chitra Seetharam
authors.initials: CS
authors.lastname: Misra
authors.affiliation: Molecular Biology Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085, India.
authors.forename: Bhakti
authors.initials: B
authors.lastname: Basu
authors.affiliation: Molecular Biology Division, Bhabha Atomic Research Centre, Trombay, Mumbai 400 085, India. Electronic address: aptesk@barc.gov.in.
authors.forename: Shree Kumar
authors.initials: SK
authors.lastname: Apte
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D002473:Cell Wall / Q000378:metabolism; D034301:Deinococcus / Q000378:metabolism*; D010457:Peptidoglycan / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013499:Surface Properties
chemical_list: D001426:Bacterial Proteins; D010457:Peptidoglycan
keywords: Deinococcus; Protein engineering; SLH domain; Surface display; Surface layer proteins