1887

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Volume 156, Issue 6

Characterization of the haem-uptake system of the equine pathogen subsp. Free

Abstract

possesses a haem-uptake system homologous to that of and The system consists of two ligand-binding proteins (Shr and Shp) and proteins (HtsA–C) with homology to an ABC transporter. The haem-uptake system of differs from that of and in that Shr is truncated by two-thirds. This study focused on the SeShr, SeShp and SeHtsA proteins of . Analysis of s, and knockout mutants showed that all three proteins were expressed and that expression was upregulated under conditions of iron limitation. SeShr possesses no membrane-/cell wall-spanning sequences and was shown to be secreted. Both SeShp and SeHtsA were confirmed to be envelope-associated. Recombinant SeShp and SeHtsA proteins have been previously shown to bind haem and SeHtsA could capture haem from SeShp. This report extends these studies and shows that both SeShp and SeHtsA can sequester haem from haemoglobin but not from haemoglobin–haptoglobin complexes. Like full-length Shr, SeShr possesses haemoglobin and haemoglobin–haptoglobin binding ability but unlike full-length Shr, it lacks haem- or fibronectin-binding capabilities. Analysis of SeShr truncates showed that residues within and upstream of the near transporter (NEAT) domain are required for this ligand binding. Structural predictions suggest that truncation of NEAT1 in SeShr accounts for its impaired ability to bind haem. Haem and haemoglobin restored to almost normal the impaired growth rates of wild-type cultured under iron-limiting conditions. However, no difference in the growth rates of wild-type and mutants could be detected under the growth conditions tested.

  • Received:
  • Accepted:
  • Revised:
  • Published Online:
Keyword(s): DIP, 2,2′ dipyridyl , Fn, fibronectin , GST, glutathione S-transferase , Hb, haemoglobin , Hp, haptoglobin , HRP, horseradish peroxidase , LB, Luria–Bertani , LRR, leucine-rich repeat , NEAT, near transporter and THYE, Todd–Hewitt yeast extract
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2010-06-01
2024-04-26
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Characterization of the haem-uptake system of the equine pathogen Streptococcus equi subsp. equi
Microbiology 156, 1824 (2010); https://doi.org/10.1099/mic.0.036087-0
/content/journal/micro/10.1099/mic.0.036087-0
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