Interactions of haemoglobin with the Neisseria meningitidis receptor HpuAB: the role of TonB and an intact proton motive force
Summary We have characterized the interaction of the Neis‐seria meningitidis TonB‐dependent receptor HpuAB with haemoglobin (Hb). Protease accessibility assays indicated that HpuA and HpuB are surface exposed, HpuB interacts physically with HpuA, and TonB energization affects the conformation of Hpu...
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| Veröffentlicht in: | Molecular microbiology 2002-01, Vol.43 (2), p.335-354 |
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| creator | Rohde, K. H. Gillaspy, A. F. Hatfield, M. D. Lewis, L. A. Dyer, D. W. |
| description | Summary
We have characterized the interaction of the Neis‐seria meningitidis TonB‐dependent receptor HpuAB with haemoglobin (Hb). Protease accessibility assays indicated that HpuA and HpuB are surface exposed, HpuB interacts physically with HpuA, and TonB energization affects the conformation of HpuAB. Binding assays using [125I]‐Hb revealed that the bipartite receptor has a single binding site for Hb (Kd≈ 150 nM). Competitive binding assays using heterologous Hbs revealed that HpuAB Hb recognition was not species specific. The binding kinetics of Hb to HpuAB were dramatically altered in a TonB– mutant and in wild‐type meningococci treated with the protonophore carbonylcyanide m‐chlorophenylhydrazone (CCCP), indicating that TonB and an intact proton motive force are required for normal Hb binding and release from HpuAB. Our results support a model in which both HpuA and HpuB are required to form a receptor complex in the outer membrane with a single binding site, whose structure and ligand interactions are significantly affected by the TonB‐mediated energy state of the receptor. |
| doi_str_mv | 10.1046/j.1365-2958.2002.02745.x |
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We have characterized the interaction of the Neis‐seria meningitidis TonB‐dependent receptor HpuAB with haemoglobin (Hb). Protease accessibility assays indicated that HpuA and HpuB are surface exposed, HpuB interacts physically with HpuA, and TonB energization affects the conformation of HpuAB. Binding assays using [125I]‐Hb revealed that the bipartite receptor has a single binding site for Hb (Kd≈ 150 nM). Competitive binding assays using heterologous Hbs revealed that HpuAB Hb recognition was not species specific. The binding kinetics of Hb to HpuAB were dramatically altered in a TonB– mutant and in wild‐type meningococci treated with the protonophore carbonylcyanide m‐chlorophenylhydrazone (CCCP), indicating that TonB and an intact proton motive force are required for normal Hb binding and release from HpuAB. Our results support a model in which both HpuA and HpuB are required to form a receptor complex in the outer membrane with a single binding site, whose structure and ligand interactions are significantly affected by the TonB‐mediated energy state of the receptor.</description><identifier>ISSN: 0950-382X</identifier><identifier>EISSN: 1365-2958</identifier><identifier>DOI: 10.1046/j.1365-2958.2002.02745.x</identifier><identifier>PMID: 11985713</identifier><language>eng</language><publisher>Oxford, UK: Blackwell Science Ltd</publisher><subject>Bacterial Outer Membrane Proteins - genetics ; Bacterial Outer Membrane Proteins - metabolism ; Bacterial Proteins - metabolism ; Hemoglobins - metabolism ; Kinetics ; Ligands ; Membrane Proteins - metabolism ; Neisseria meningitidis - genetics ; Neisseria meningitidis - growth & development ; Neisseria meningitidis - metabolism ; Receptors, Cell Surface - genetics ; Receptors, Cell Surface - metabolism ; Trypsin - metabolism</subject><ispartof>Molecular microbiology, 2002-01, Vol.43 (2), p.335-354</ispartof><rights>Copyright Blackwell Scientific Publications Ltd. Jan 2002</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4735-3819e3a2f8f634a00e1c24b264d4112e23efd0e9b92d81790aaa38463ba627e03</citedby><cites>FETCH-LOGICAL-c4735-3819e3a2f8f634a00e1c24b264d4112e23efd0e9b92d81790aaa38463ba627e03</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1046%2Fj.1365-2958.2002.02745.x$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1046%2Fj.1365-2958.2002.02745.x$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,777,781,1412,1428,27905,27906,45555,45556,46390,46814</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/11985713$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Rohde, K. H.</creatorcontrib><creatorcontrib>Gillaspy, A. F.</creatorcontrib><creatorcontrib>Hatfield, M. D.</creatorcontrib><creatorcontrib>Lewis, L. A.</creatorcontrib><creatorcontrib>Dyer, D. W.</creatorcontrib><title>Interactions of haemoglobin with the Neisseria meningitidis receptor HpuAB: the role of TonB and an intact proton motive force</title><title>Molecular microbiology</title><addtitle>Mol Microbiol</addtitle><description>Summary
We have characterized the interaction of the Neis‐seria meningitidis TonB‐dependent receptor HpuAB with haemoglobin (Hb). Protease accessibility assays indicated that HpuA and HpuB are surface exposed, HpuB interacts physically with HpuA, and TonB energization affects the conformation of HpuAB. Binding assays using [125I]‐Hb revealed that the bipartite receptor has a single binding site for Hb (Kd≈ 150 nM). Competitive binding assays using heterologous Hbs revealed that HpuAB Hb recognition was not species specific. The binding kinetics of Hb to HpuAB were dramatically altered in a TonB– mutant and in wild‐type meningococci treated with the protonophore carbonylcyanide m‐chlorophenylhydrazone (CCCP), indicating that TonB and an intact proton motive force are required for normal Hb binding and release from HpuAB. Our results support a model in which both HpuA and HpuB are required to form a receptor complex in the outer membrane with a single binding site, whose structure and ligand interactions are significantly affected by the TonB‐mediated energy state of the receptor.</description><subject>Bacterial Outer Membrane Proteins - genetics</subject><subject>Bacterial Outer Membrane Proteins - metabolism</subject><subject>Bacterial Proteins - metabolism</subject><subject>Hemoglobins - metabolism</subject><subject>Kinetics</subject><subject>Ligands</subject><subject>Membrane Proteins - metabolism</subject><subject>Neisseria meningitidis - genetics</subject><subject>Neisseria meningitidis - growth & development</subject><subject>Neisseria meningitidis - metabolism</subject><subject>Receptors, Cell Surface - genetics</subject><subject>Receptors, Cell Surface - metabolism</subject><subject>Trypsin - metabolism</subject><issn>0950-382X</issn><issn>1365-2958</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2002</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqNkU1v1DAQhi1ERZfSv4AsDtw29VdsB4lDWwFdqYVLkXqznGTS9SqxF9vpx4XfTtJdtRIXOFhjaZ557dGDEKakoETIk01BuSyXrCp1wQhhBWFKlMXDK7R4brxGC1KVZMk1uzlEb1PaEEI5kfwNOqS00qWifIF-r3yGaJvsgk84dHhtYQi3faidx_cur3FeA_4OLiWIzuIBvPO3LrvWJRyhgW0OEV9sx9OzT09oDD3MOdfBn2Hr2-lg5_P0At7GkIPHQ8juDnAXYgPv0EFn-wTH-3qEfn79cn1-sbz88W11fnq5bITi5bQDrYBb1ulOcmEJAdowUTMpWkEpA8ahawlUdcVaTVVFrLVcC8lrK5kCwo_Qx13u9IdfI6RsBpca6HvrIYzJKCpVKQX_J0i1kloQMYEf_gI3YYx-WsLQSpZEsYpOkN5BTQwpRejMNrrBxkdDiZlNmo2ZhZlZmJlNmieT5mEafb_PH-sB2pfBvboJ-LwD7l0Pj_8dbK6uVvON_wEO26yi</recordid><startdate>200201</startdate><enddate>200201</enddate><creator>Rohde, K. H.</creator><creator>Gillaspy, A. F.</creator><creator>Hatfield, M. D.</creator><creator>Lewis, L. A.</creator><creator>Dyer, D. W.</creator><general>Blackwell Science Ltd</general><general>Blackwell Publishing Ltd</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7TK</scope><scope>7TM</scope><scope>7U9</scope><scope>8FD</scope><scope>C1K</scope><scope>FR3</scope><scope>H94</scope><scope>M7N</scope><scope>P64</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>200201</creationdate><title>Interactions of haemoglobin with the Neisseria meningitidis receptor HpuAB: the role of TonB and an intact proton motive force</title><author>Rohde, K. H. ; Gillaspy, A. F. ; Hatfield, M. D. ; Lewis, L. A. ; Dyer, D. W.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4735-3819e3a2f8f634a00e1c24b264d4112e23efd0e9b92d81790aaa38463ba627e03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2002</creationdate><topic>Bacterial Outer Membrane Proteins - genetics</topic><topic>Bacterial Outer Membrane Proteins - metabolism</topic><topic>Bacterial Proteins - metabolism</topic><topic>Hemoglobins - metabolism</topic><topic>Kinetics</topic><topic>Ligands</topic><topic>Membrane Proteins - metabolism</topic><topic>Neisseria meningitidis - genetics</topic><topic>Neisseria meningitidis - growth & development</topic><topic>Neisseria meningitidis - metabolism</topic><topic>Receptors, Cell Surface - genetics</topic><topic>Receptors, Cell Surface - metabolism</topic><topic>Trypsin - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Rohde, K. H.</creatorcontrib><creatorcontrib>Gillaspy, A. F.</creatorcontrib><creatorcontrib>Hatfield, M. D.</creatorcontrib><creatorcontrib>Lewis, L. A.</creatorcontrib><creatorcontrib>Dyer, D. W.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Algology Mycology and Protozoology Abstracts (Microbiology C)</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Molecular microbiology</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Rohde, K. H.</au><au>Gillaspy, A. F.</au><au>Hatfield, M. D.</au><au>Lewis, L. A.</au><au>Dyer, D. W.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Interactions of haemoglobin with the Neisseria meningitidis receptor HpuAB: the role of TonB and an intact proton motive force</atitle><jtitle>Molecular microbiology</jtitle><addtitle>Mol Microbiol</addtitle><date>2002-01</date><risdate>2002</risdate><volume>43</volume><issue>2</issue><spage>335</spage><epage>354</epage><pages>335-354</pages><issn>0950-382X</issn><eissn>1365-2958</eissn><abstract>Summary
We have characterized the interaction of the Neis‐seria meningitidis TonB‐dependent receptor HpuAB with haemoglobin (Hb). Protease accessibility assays indicated that HpuA and HpuB are surface exposed, HpuB interacts physically with HpuA, and TonB energization affects the conformation of HpuAB. Binding assays using [125I]‐Hb revealed that the bipartite receptor has a single binding site for Hb (Kd≈ 150 nM). Competitive binding assays using heterologous Hbs revealed that HpuAB Hb recognition was not species specific. The binding kinetics of Hb to HpuAB were dramatically altered in a TonB– mutant and in wild‐type meningococci treated with the protonophore carbonylcyanide m‐chlorophenylhydrazone (CCCP), indicating that TonB and an intact proton motive force are required for normal Hb binding and release from HpuAB. Our results support a model in which both HpuA and HpuB are required to form a receptor complex in the outer membrane with a single binding site, whose structure and ligand interactions are significantly affected by the TonB‐mediated energy state of the receptor.</abstract><cop>Oxford, UK</cop><pub>Blackwell Science Ltd</pub><pmid>11985713</pmid><doi>10.1046/j.1365-2958.2002.02745.x</doi><tpages>20</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Bacterial Outer Membrane Proteins - genetics Bacterial Outer Membrane Proteins - metabolism Bacterial Proteins - metabolism Hemoglobins - metabolism Kinetics Ligands Membrane Proteins - metabolism Neisseria meningitidis - genetics Neisseria meningitidis - growth & development Neisseria meningitidis - metabolism Receptors, Cell Surface - genetics Receptors, Cell Surface - metabolism Trypsin - metabolism |
| title | Interactions of haemoglobin with the Neisseria meningitidis receptor HpuAB: the role of TonB and an intact proton motive force |
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