Restoring Allosterism with Compensatory Mutations in Hemoglobin
Abnormal human hemoglobins (Hbs) with amino acid substitutions in the α1β2interface have very high oxygen affinity and greatly reduce cooperativity in O2binding compared to normal human Hb. In such abnormal Hbs with mutations at position β99, the intersubunit hydrogen bonds between Asp-β99 and Tyr-α...
Gespeichert in:
| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1994-11, Vol.91 (24), p.11547-11551 |
|---|---|
| Hauptverfasser: | , , , , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Schlagworte: | |
| Online-Zugang: | Volltext |
| Tags: |
Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
|
| container_end_page | 11551 |
|---|---|
| container_issue | 24 |
| container_start_page | 11547 |
| container_title | Proceedings of the National Academy of Sciences - PNAS |
| container_volume | 91 |
| creator | Kim, H W Shen, T J Sun, D P Ho, N T Madrid, M Tam, M F Zou, M Cottam, P F Ho, C |
| description | Abnormal human hemoglobins (Hbs) with amino acid substitutions in the α1β2interface have very high oxygen affinity and greatly reduce cooperativity in O2binding compared to normal human Hb. In such abnormal Hbs with mutations at position β99, the intersubunit hydrogen bonds between Asp-β99 and Tyr-α42 and between Asp- β99 and Asn-α97 are broken, thus destabilizing the deoxyquaternary structure of these Hbs. A molecular dynamics method has been used to design compensatory amino acid substitutions in these Hbs that can restore their allosteric properties. We have designed a compensatory mutation in a naturally occuring mutant Hb, Hb Kempsey (Asp- β99 → Asn), and have produced it using our Escherichia coli expression plasmid pHE2. We have determined the O2binding properties of this recombinant double mutant Hb, Hb(Asp-β99 → Asn and Tyr-α42 → Asp) and have used1NMR spectroscopy to investigate the tertiary structures around the heme groups and the quaternary structure in the α1β2subunit interface. Our results clearly show that the Tyr-α42 → Asp replacement can substantially compensate for the functional defect of Hb Kempsey caused by the Asp-β99 → Asn substitution. The structural and functional information derived from this recombinant Hb provides insights into the structural basis of allosterism and the design of compensatory amino acid substitutions to restore the functional properties of other abnormal Hbs associated with hemoglobinopathies. |
| doi_str_mv | 10.1073/pnas.91.24.11547 |
| format | Article |
| fullrecord | <record><control><sourceid>jstor_pubme</sourceid><recordid>TN_cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_45268</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><jstor_id>2366167</jstor_id><sourcerecordid>2366167</sourcerecordid><originalsourceid>FETCH-LOGICAL-c525t-da99ea9b5438669cb4f8a269222497882fbdff789dfc8d0a2fe463f4a7ac58e03</originalsourceid><addsrcrecordid>eNp9kc9rFDEYhoNY6lq9e1AcPEgvsyaZ_ARByqKtUBFEzyEzk2yzZJI1yaj9782626V68PQd3uf5-D5eAJ4huESQd2-2QeelREtMlghRwh-ABYIStYxI-BAsIMS8FQSTR-BxzhsIoaQCnoJTLjmGUi7Auy8ml5hcWDcX3sdcTHJ5an66ctOs4rQ1Ieua3zaf5qKLiyE3LjRXZoprH3sXnoATq302Tw_zDHz78P7r6qq9_nz5cXVx3Q4U09KOWkqjZU9JJxiTQ0-s0JhJjDGRXAhs-9FaLuRoBzFCja0hrLNEcz1QYWB3Bt7u927nfjLjYEJJ2qttcpNOtypqp_5OgrtR6_hDEYqZqPrrg57i97m-rCaXB-O9DibOWSFGuYBIVvDVP-AmzinU1xSGCAuGKaoQ3ENDijknY493IKh2vahdL0oihYn600tVXty__ygciqj5-SHfmXfpvQ3Kzt4X86tU9OX_0Uo83xObXbdHBHeMIca734qErNM</addsrcrecordid><sourcetype>Open Access Repository</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>201286251</pqid></control><display><type>article</type><title>Restoring Allosterism with Compensatory Mutations in Hemoglobin</title><source>MEDLINE</source><source>JSTOR Archive Collection A-Z Listing</source><source>PubMed Central</source><source>Alma/SFX Local Collection</source><source>Free Full-Text Journals in Chemistry</source><creator>Kim, H W ; Shen, T J ; Sun, D P ; Ho, N T ; Madrid, M ; Tam, M F ; Zou, M ; Cottam, P F ; Ho, C</creator><creatorcontrib>Kim, H W ; Shen, T J ; Sun, D P ; Ho, N T ; Madrid, M ; Tam, M F ; Zou, M ; Cottam, P F ; Ho, C</creatorcontrib><description>Abnormal human hemoglobins (Hbs) with amino acid substitutions in the α1β2interface have very high oxygen affinity and greatly reduce cooperativity in O2binding compared to normal human Hb. In such abnormal Hbs with mutations at position β99, the intersubunit hydrogen bonds between Asp-β99 and Tyr-α42 and between Asp- β99 and Asn-α97 are broken, thus destabilizing the deoxyquaternary structure of these Hbs. A molecular dynamics method has been used to design compensatory amino acid substitutions in these Hbs that can restore their allosteric properties. We have designed a compensatory mutation in a naturally occuring mutant Hb, Hb Kempsey (Asp- β99 → Asn), and have produced it using our Escherichia coli expression plasmid pHE2. We have determined the O2binding properties of this recombinant double mutant Hb, Hb(Asp-β99 → Asn and Tyr-α42 → Asp) and have used1NMR spectroscopy to investigate the tertiary structures around the heme groups and the quaternary structure in the α1β2subunit interface. Our results clearly show that the Tyr-α42 → Asp replacement can substantially compensate for the functional defect of Hb Kempsey caused by the Asp-β99 → Asn substitution. The structural and functional information derived from this recombinant Hb provides insights into the structural basis of allosterism and the design of compensatory amino acid substitutions to restore the functional properties of other abnormal Hbs associated with hemoglobinopathies.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.91.24.11547</identifier><identifier>PMID: 7972099</identifier><language>eng</language><publisher>United States: National Academy of Sciences of the United States of America</publisher><subject>allosteric properties ; Allosteric Regulation ; Amino acid substitution ; Amino acids ; Base Sequence ; Biochemistry ; Computer Simulation ; DNA Primers - chemistry ; Globins - chemistry ; Hemoglobin ; Hemoglobins ; Hemoglobins - chemistry ; Humans ; Hydrogen bonds ; Magnetic Resonance Spectroscopy ; man ; Models, Molecular ; Molecular Sequence Data ; Mutagenesis, Site-Directed ; Mutation ; Oxygen ; Oxyhemoglobins - chemistry ; Phosphates ; Plasmids ; Protein Conformation ; Protons ; Sodium ; Structure-Activity Relationship ; Tissue oxygenation</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 1994-11, Vol.91 (24), p.11547-11551</ispartof><rights>Copyright 1994 The National Academy of Sciences of the United States of America</rights><rights>Copyright National Academy of Sciences Nov 22, 1994</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c525t-da99ea9b5438669cb4f8a269222497882fbdff789dfc8d0a2fe463f4a7ac58e03</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/91/24.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/2366167$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/2366167$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,727,780,784,803,885,27924,27925,53791,53793,58017,58250</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/7972099$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kim, H W</creatorcontrib><creatorcontrib>Shen, T J</creatorcontrib><creatorcontrib>Sun, D P</creatorcontrib><creatorcontrib>Ho, N T</creatorcontrib><creatorcontrib>Madrid, M</creatorcontrib><creatorcontrib>Tam, M F</creatorcontrib><creatorcontrib>Zou, M</creatorcontrib><creatorcontrib>Cottam, P F</creatorcontrib><creatorcontrib>Ho, C</creatorcontrib><title>Restoring Allosterism with Compensatory Mutations in Hemoglobin</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>Abnormal human hemoglobins (Hbs) with amino acid substitutions in the α1β2interface have very high oxygen affinity and greatly reduce cooperativity in O2binding compared to normal human Hb. In such abnormal Hbs with mutations at position β99, the intersubunit hydrogen bonds between Asp-β99 and Tyr-α42 and between Asp- β99 and Asn-α97 are broken, thus destabilizing the deoxyquaternary structure of these Hbs. A molecular dynamics method has been used to design compensatory amino acid substitutions in these Hbs that can restore their allosteric properties. We have designed a compensatory mutation in a naturally occuring mutant Hb, Hb Kempsey (Asp- β99 → Asn), and have produced it using our Escherichia coli expression plasmid pHE2. We have determined the O2binding properties of this recombinant double mutant Hb, Hb(Asp-β99 → Asn and Tyr-α42 → Asp) and have used1NMR spectroscopy to investigate the tertiary structures around the heme groups and the quaternary structure in the α1β2subunit interface. Our results clearly show that the Tyr-α42 → Asp replacement can substantially compensate for the functional defect of Hb Kempsey caused by the Asp-β99 → Asn substitution. The structural and functional information derived from this recombinant Hb provides insights into the structural basis of allosterism and the design of compensatory amino acid substitutions to restore the functional properties of other abnormal Hbs associated with hemoglobinopathies.</description><subject>allosteric properties</subject><subject>Allosteric Regulation</subject><subject>Amino acid substitution</subject><subject>Amino acids</subject><subject>Base Sequence</subject><subject>Biochemistry</subject><subject>Computer Simulation</subject><subject>DNA Primers - chemistry</subject><subject>Globins - chemistry</subject><subject>Hemoglobin</subject><subject>Hemoglobins</subject><subject>Hemoglobins - chemistry</subject><subject>Humans</subject><subject>Hydrogen bonds</subject><subject>Magnetic Resonance Spectroscopy</subject><subject>man</subject><subject>Models, Molecular</subject><subject>Molecular Sequence Data</subject><subject>Mutagenesis, Site-Directed</subject><subject>Mutation</subject><subject>Oxygen</subject><subject>Oxyhemoglobins - chemistry</subject><subject>Phosphates</subject><subject>Plasmids</subject><subject>Protein Conformation</subject><subject>Protons</subject><subject>Sodium</subject><subject>Structure-Activity Relationship</subject><subject>Tissue oxygenation</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1994</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kc9rFDEYhoNY6lq9e1AcPEgvsyaZ_ARByqKtUBFEzyEzk2yzZJI1yaj9782626V68PQd3uf5-D5eAJ4huESQd2-2QeelREtMlghRwh-ABYIStYxI-BAsIMS8FQSTR-BxzhsIoaQCnoJTLjmGUi7Auy8ml5hcWDcX3sdcTHJ5an66ctOs4rQ1Ieua3zaf5qKLiyE3LjRXZoprH3sXnoATq302Tw_zDHz78P7r6qq9_nz5cXVx3Q4U09KOWkqjZU9JJxiTQ0-s0JhJjDGRXAhs-9FaLuRoBzFCja0hrLNEcz1QYWB3Bt7u927nfjLjYEJJ2qttcpNOtypqp_5OgrtR6_hDEYqZqPrrg57i97m-rCaXB-O9DibOWSFGuYBIVvDVP-AmzinU1xSGCAuGKaoQ3ENDijknY493IKh2vahdL0oihYn600tVXty__ygciqj5-SHfmXfpvQ3Kzt4X86tU9OX_0Uo83xObXbdHBHeMIca734qErNM</recordid><startdate>19941122</startdate><enddate>19941122</enddate><creator>Kim, H W</creator><creator>Shen, T J</creator><creator>Sun, D P</creator><creator>Ho, N T</creator><creator>Madrid, M</creator><creator>Tam, M F</creator><creator>Zou, M</creator><creator>Cottam, P F</creator><creator>Ho, C</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</general><general>National Academy of Sciences</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>7QG</scope><scope>7QL</scope><scope>7QP</scope><scope>7QR</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TK</scope><scope>7TM</scope><scope>7TO</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>7T3</scope><scope>5PM</scope></search><sort><creationdate>19941122</creationdate><title>Restoring Allosterism with Compensatory Mutations in Hemoglobin</title><author>Kim, H W ; Shen, T J ; Sun, D P ; Ho, N T ; Madrid, M ; Tam, M F ; Zou, M ; Cottam, P F ; Ho, C</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c525t-da99ea9b5438669cb4f8a269222497882fbdff789dfc8d0a2fe463f4a7ac58e03</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1994</creationdate><topic>allosteric properties</topic><topic>Allosteric Regulation</topic><topic>Amino acid substitution</topic><topic>Amino acids</topic><topic>Base Sequence</topic><topic>Biochemistry</topic><topic>Computer Simulation</topic><topic>DNA Primers - chemistry</topic><topic>Globins - chemistry</topic><topic>Hemoglobin</topic><topic>Hemoglobins</topic><topic>Hemoglobins - chemistry</topic><topic>Humans</topic><topic>Hydrogen bonds</topic><topic>Magnetic Resonance Spectroscopy</topic><topic>man</topic><topic>Models, Molecular</topic><topic>Molecular Sequence Data</topic><topic>Mutagenesis, Site-Directed</topic><topic>Mutation</topic><topic>Oxygen</topic><topic>Oxyhemoglobins - chemistry</topic><topic>Phosphates</topic><topic>Plasmids</topic><topic>Protein Conformation</topic><topic>Protons</topic><topic>Sodium</topic><topic>Structure-Activity Relationship</topic><topic>Tissue oxygenation</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kim, H W</creatorcontrib><creatorcontrib>Shen, T J</creatorcontrib><creatorcontrib>Sun, D P</creatorcontrib><creatorcontrib>Ho, N T</creatorcontrib><creatorcontrib>Madrid, M</creatorcontrib><creatorcontrib>Tam, M F</creatorcontrib><creatorcontrib>Zou, M</creatorcontrib><creatorcontrib>Cottam, P F</creatorcontrib><creatorcontrib>Ho, C</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Animal Behavior Abstracts</collection><collection>Bacteriology Abstracts (Microbiology B)</collection><collection>Calcium & Calcified Tissue Abstracts</collection><collection>Chemoreception Abstracts</collection><collection>Ecology Abstracts</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Immunology Abstracts</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Oncogenes and Growth Factors 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>Human Genome Abstracts</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kim, H W</au><au>Shen, T J</au><au>Sun, D P</au><au>Ho, N T</au><au>Madrid, M</au><au>Tam, M F</au><au>Zou, M</au><au>Cottam, P F</au><au>Ho, C</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Restoring Allosterism with Compensatory Mutations in Hemoglobin</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1994-11-22</date><risdate>1994</risdate><volume>91</volume><issue>24</issue><spage>11547</spage><epage>11551</epage><pages>11547-11551</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>Abnormal human hemoglobins (Hbs) with amino acid substitutions in the α1β2interface have very high oxygen affinity and greatly reduce cooperativity in O2binding compared to normal human Hb. In such abnormal Hbs with mutations at position β99, the intersubunit hydrogen bonds between Asp-β99 and Tyr-α42 and between Asp- β99 and Asn-α97 are broken, thus destabilizing the deoxyquaternary structure of these Hbs. A molecular dynamics method has been used to design compensatory amino acid substitutions in these Hbs that can restore their allosteric properties. We have designed a compensatory mutation in a naturally occuring mutant Hb, Hb Kempsey (Asp- β99 → Asn), and have produced it using our Escherichia coli expression plasmid pHE2. We have determined the O2binding properties of this recombinant double mutant Hb, Hb(Asp-β99 → Asn and Tyr-α42 → Asp) and have used1NMR spectroscopy to investigate the tertiary structures around the heme groups and the quaternary structure in the α1β2subunit interface. Our results clearly show that the Tyr-α42 → Asp replacement can substantially compensate for the functional defect of Hb Kempsey caused by the Asp-β99 → Asn substitution. The structural and functional information derived from this recombinant Hb provides insights into the structural basis of allosterism and the design of compensatory amino acid substitutions to restore the functional properties of other abnormal Hbs associated with hemoglobinopathies.</abstract><cop>United States</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>7972099</pmid><doi>10.1073/pnas.91.24.11547</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 0027-8424 |
| ispartof | Proceedings of the National Academy of Sciences - PNAS, 1994-11, Vol.91 (24), p.11547-11551 |
| issn | 0027-8424 1091-6490 |
| language | eng |
| recordid | cdi_pubmedcentral_primary_oai_pubmedcentral_nih_gov_45268 |
| source | MEDLINE; JSTOR Archive Collection A-Z Listing; PubMed Central; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry |
| subjects | allosteric properties Allosteric Regulation Amino acid substitution Amino acids Base Sequence Biochemistry Computer Simulation DNA Primers - chemistry Globins - chemistry Hemoglobin Hemoglobins Hemoglobins - chemistry Humans Hydrogen bonds Magnetic Resonance Spectroscopy man Models, Molecular Molecular Sequence Data Mutagenesis, Site-Directed Mutation Oxygen Oxyhemoglobins - chemistry Phosphates Plasmids Protein Conformation Protons Sodium Structure-Activity Relationship Tissue oxygenation |
| title | Restoring Allosterism with Compensatory Mutations in Hemoglobin |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-21T17%3A51%3A26IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-jstor_pubme&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Restoring%20Allosterism%20with%20Compensatory%20Mutations%20in%20Hemoglobin&rft.jtitle=Proceedings%20of%20the%20National%20Academy%20of%20Sciences%20-%20PNAS&rft.au=Kim,%20H%20W&rft.date=1994-11-22&rft.volume=91&rft.issue=24&rft.spage=11547&rft.epage=11551&rft.pages=11547-11551&rft.issn=0027-8424&rft.eissn=1091-6490&rft_id=info:doi/10.1073/pnas.91.24.11547&rft_dat=%3Cjstor_pubme%3E2366167%3C/jstor_pubme%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=201286251&rft_id=info:pmid/7972099&rft_jstor_id=2366167&rfr_iscdi=true |