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dc.contributor.authorJohnson, Eric A.
dc.contributor.authorRice, Selena L.
dc.contributor.authorPreimesberger, Matthew R.
dc.contributor.authorNye, Dillon B.
dc.contributor.authorGilevicius, Lukas
dc.contributor.authorWenke, Belinda B.
dc.contributor.authorBrown, Jason
dc.contributor.authorWitman, George B.
dc.contributor.authorLecomte, Juliette T. J.
dc.date2022-08-11T08:08:03.000
dc.date.accessioned2022-08-23T15:40:42Z
dc.date.available2022-08-23T15:40:42Z
dc.date.issued2014-07-22
dc.date.submitted2015-04-01
dc.identifier.citation<p>Biochemistry. 2014 Jul 22;53(28):4573-89. doi: 10.1021/bi5005206. <a href="http://dx.doi.org/10.1021/bi5005206">Link to article on publisher's site</a>.</p>
dc.identifier.issn0006-2960 (Linking)
dc.identifier.doi10.1021/bi5005206
dc.identifier.pmid24964018
dc.identifier.urihttp://hdl.handle.net/20.500.14038/26463
dc.description.abstractThe nuclear genome of the model organism Chlamydomonas reinhardtii contains genes for a dozen hemoglobins of the truncated lineage. Of those, THB1 is known to be expressed, but the product and its function have not yet been characterized. We present mutagenesis, optical, and nuclear magnetic resonance data for the recombinant protein and show that at pH near neutral in the absence of added ligand, THB1 coordinates the heme iron with the canonical proximal histidine and a distal lysine. In the cyanomet state, THB1 is structurally similar to other known truncated hemoglobins, particularly the heme domain of Chlamydomonas eugametos LI637, a light-induced chloroplastic hemoglobin. Recombinant THB1 is capable of binding nitric oxide (NO(*)) in either the ferric or ferrous state and has efficient NO(*) dioxygenase activity. By using different C. reinhardtii strains and growth conditions, we demonstrate that the expression of THB1 is under the control of the NIT2 regulatory gene and that the hemoglobin is linked to the nitrogen assimilation pathway.
dc.language.isoen_US
dc.relation<p><a href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=pubmed&cmd=Retrieve&list_uids=24964018&dopt=Abstract">Link to Article in PubMed</a></p>
dc.rights<p>Open access via the <a href="http://pubs.acs.org/page/policy/authorchoice/index.html" target="_blank" title="ACS AuthorChoice">ACS AuthorChoice + 12, Open Access</a> option on 06/25/2015.</p>
dc.subjectChlamydomonas reinhardtii
dc.subjectChloroplast Proteins
dc.subjectGene Expression Regulation, Plant
dc.subjectHeme
dc.subjectHemoglobins
dc.subjectHydrogen-Ion Concentration
dc.subjectLysine
dc.subjectNitric Oxide
dc.subjectNitrogen
dc.subjectAlgae
dc.subjectAmino Acids, Peptides, and Proteins
dc.subjectBiochemical Phenomena, Metabolism, and Nutrition
dc.subjectBiochemistry
dc.subjectChemical Actions and Uses
dc.subjectGenetic Phenomena
dc.subjectInorganic Chemicals
dc.subjectInvestigative Techniques
dc.titleCharacterization of THB1, a Chlamydomonas reinhardtii truncated hemoglobin: linkage to nitrogen metabolism and identification of lysine as the distal heme ligand
dc.typeJournal Article
dc.source.journaltitleBiochemistry
dc.source.volume53
dc.source.issue28
dc.identifier.legacyfulltexthttps://escholarship.umassmed.edu/cgi/viewcontent.cgi?article=1147&amp;context=cellbiology_pp&amp;unstamped=1
dc.identifier.legacycoverpagehttps://escholarship.umassmed.edu/cellbiology_pp/148
dc.legacy.embargo2015-06-25T00:00:00-07:00
dc.identifier.contextkey6929798
refterms.dateFOA2022-08-23T15:40:43Z
html.description.abstract<p>The nuclear genome of the model organism Chlamydomonas reinhardtii contains genes for a dozen hemoglobins of the truncated lineage. Of those, THB1 is known to be expressed, but the product and its function have not yet been characterized. We present mutagenesis, optical, and nuclear magnetic resonance data for the recombinant protein and show that at pH near neutral in the absence of added ligand, THB1 coordinates the heme iron with the canonical proximal histidine and a distal lysine. In the cyanomet state, THB1 is structurally similar to other known truncated hemoglobins, particularly the heme domain of Chlamydomonas eugametos LI637, a light-induced chloroplastic hemoglobin. Recombinant THB1 is capable of binding nitric oxide (NO(*)) in either the ferric or ferrous state and has efficient NO(*) dioxygenase activity. By using different C. reinhardtii strains and growth conditions, we demonstrate that the expression of THB1 is under the control of the NIT2 regulatory gene and that the hemoglobin is linked to the nitrogen assimilation pathway.</p>
dc.identifier.submissionpathcellbiology_pp/148
dc.contributor.departmentDepartment of Cell and Developmental Biology
dc.source.pages4573-89


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