The Streptococcus pyogenes Shr protein captures human hemoglobin using two structurally unique binding domains

Macdonald, Ramsay; Cascio, Duilio; Collazo, Michael J.; Phillips, Martin; Clubb, Robert T.

doi:10.1074/jbc.ra118.005261

Title: The Streptococcus pyogenes Shr protein captures human hemoglobin using two structurally unique binding domains

Journal Article · Tue Oct 09 00:00:00 EDT 2018 · Journal of Biological Chemistry

DOI:https://doi.org/10.1074/jbc.ra118.005261· OSTI ID:1560082

Macdonald, Ramsay ^[1]; Cascio, Duilio ^[1]; Collazo, Michael J. ^[1]; Phillips, Martin ^[1]; Clubb, Robert T. ^[1]

Univ. of California, Los Angeles, CA (United States)

In order to proliferate and mount an infection, many bacterial pathogens need to acquire iron from their host. The most abundant iron source in the body is the oxygen transporter hemoglobin (Hb). Streptococcus pyogenes, a potentially lethal human pathogen, uses the Shr protein to capture Hb on the cell surface. Shr is an important virulence factor, yet the mechanism by which it captures Hb and acquires its heme is not well-understood. In this work, we show using NMR and biochemical methods that Shr binds Hb using two related modules that were previously defined as domains of unknown function (DUF1533). These hemoglobin-interacting domains (HIDs), called HID1 and HID2, are autonomously folded and independently bind Hb. The 1.5 Å resolution crystal structure of HID2 revealed that it is a structurally unique Hb-binding domain. Mutagenesis studies revealed a conserved tyrosine in both HIDs that is essential for Hb binding. Our biochemical studies indicate that HID2 binds Hb with higher affinity than HID1 and that the Hb tetramer is engaged by two Shr receptors. NMR studies reveal the presence of a third autonomously folded domain between HID2 and a heme-binding NEAT1 domain, suggesting that this linker domain may position NEAT1 near Hb for heme capture.

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Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States). Advanced Photon Source (APS)

Sponsoring Organization:: National Institutes of Health (NIH); National Institute of General Medical Sciences (NIGMS); Office of Research Infrastructure Programs (ORIP); USDOE Office of Science (SC); National Science Foundation (NSF)

Grant/Contract Number:: S10OD016336; P41 GM103403; S10 RR029205; AC02-06CH11357; MCB-1716948; AI052217

OSTI ID:: 1560082

Journal Information:: Journal of Biological Chemistry, Vol. 293, Issue 47; ISSN 0021-9258

Publisher:: American Society for Biochemistry and Molecular BiologyCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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Cited by: 5 works

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59 BASIC BIOLOGICAL SCIENCES
37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY
Streptococcus pyogenes (S. pyogenes)
hemoglobin
X-ray crystallography
nuclear magnetic resonance (NMR)
isothermal titration calorimetry (ITC)
receptor
bacterial pathogen
DUF1533
Shr

Title: The Streptococcus pyogenes Shr protein captures human hemoglobin using two structurally unique binding domains

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