Identification and characterization of a <i>Streptococcus pyogenes</i> ABC transporter with multiple specificity for metal cations

Molecular Microbiology - Tập 34 Số 3 - Trang 596-606 - 1999
Robert Janulczyk1, Jan Pallon2, Lars Björck1
1Department of Cell and Molecular Biology, Section for Molecular Pathogenesis, and the,
2Department of Nuclear Physics, Lund University, PO Box 94, S-221 00 Lund, Sweden.

Tóm tắt

Metal ions are crucial trace elements for bacteria infecting the human host. The LraI (lipoprotein receptor‐associated antigen I) transporter in Streptococcus spp. belongs to the superfamily of ABC transporters. The transporter consists of a lipoprotein, an ATP‐binding protein and a hydrophobic integral membrane protein. Here, we describe a new member of the LraI family in the important human pathogen Streptococcus pyogenes. The system was identified in silico by analysis of the S. pyogenes Genome Sequencing Project. The S. pyogenes operon exhibits an atypical organization compared with equivalents in other Streptococcus spp. The presence and atypical organization of the operon was verified in a number of S. pyogenes strains of different serotypes. Transcriptional analysis of the LraI operon demonstrates a polycistronic transcription attenuated by a stable stem–loop structure, which allows the lipoprotein to be expressed in larger quantities than the other two components. The localization of the native lipoprotein at the bacterial surface was shown by proteolytic digestion of S. pyogenes bacteria and NH2‐terminal sequencing of a released lipoprotein fragment. Recombinant lipoprotein was expressed as a GST fusion protein, and studies of molecular interactions with metal radioisotopes demonstrated that the protein has affinity for Zn(II), Fe(III) and Cu(II). Zn(II) and Cu(II) were found to compete for the same binding site, whereas Fe(III) uses a second site. Also, proton‐induced X‐ray analysis of lipoprotein samples identified iron, copper and zinc. Finally, a mutant strain lacking a functional mtsABC operon was generated and showed reduced uptake of 55Fe and 65Zn compared with the wild‐type strain. The operon encoding this novel ABC transporter with multiple specificity for metal cations is designated mtsABC, for metal transporter of Streptococcus.

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Molecular Microbiology

Tập 34 Số 3

596-606

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