Transient B12-Dependent Methyltransferase Complexes Revealed by Small-Angle X-ray Scattering
In the Wood−Ljungdahl carbon fixation pathway, protein−protein interactions between methyltransferase (MeTr) and corrinoid iron−sulfur protein (CFeSP) are required for the transfer of a methyl group. While crystal structures have been determined for MeTr and CFeSP both free and in complex, solution...
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| Format: | Article |
| Language: | en_US |
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American Chemical Society
2013
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| Online Access: | http://hdl.handle.net/1721.1/82017 https://orcid.org/0000-0001-5486-2755 |
| _version_ | 1811078708854784000 |
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| author | Ando, Nozomi Kung, Yan Can, Mehmet Bender, Güneş Ragsdale, Stephen W. Drennan, Catherine L |
| author2 | Massachusetts Institute of Technology. Department of Biology |
| author_facet | Massachusetts Institute of Technology. Department of Biology Ando, Nozomi Kung, Yan Can, Mehmet Bender, Güneş Ragsdale, Stephen W. Drennan, Catherine L |
| author_sort | Ando, Nozomi |
| collection | MIT |
| description | In the Wood−Ljungdahl carbon fixation pathway, protein−protein interactions between methyltransferase (MeTr) and corrinoid iron−sulfur protein (CFeSP) are required for the transfer of a methyl group. While crystal structures have been determined for MeTr and CFeSP both free and in complex, solution structures have not been established. Here, we examine the transient interactions between MeTr and CFeSP in solution using anaerobic small-angle Xray scattering (SAXS) and present a global analysis approach for the deconvolution of heterogeneous mixtures formed by weakly interacting proteins. We further support this SAXS analysis with complementary results obtained by anaerobic isothermal titration calorimetry. Our results indicate that solution conditions affect the cooperativity with which CFeSP binds to MeTr, resulting in two distinct CFeSP/MeTr complexes with differing oligomeric compositions, both of which are active. One assembly resembles the CFeSP/MeTr complex observed crystallographically with 2:1 protein stoichiometry, while the other best fits a 1:1 CFeSP/MeTr arrangement. These results demonstrate the value of SAXS in uncovering the rich solution behavior of transient protein interactions visualized by crystallography. |
| first_indexed | 2024-09-23T11:04:12Z |
| format | Article |
| id | mit-1721.1/82017 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T11:04:12Z |
| publishDate | 2013 |
| publisher | American Chemical Society |
| record_format | dspace |
| spelling | mit-1721.1/820172022-09-27T16:54:48Z Transient B12-Dependent Methyltransferase Complexes Revealed by Small-Angle X-ray Scattering Ando, Nozomi Kung, Yan Can, Mehmet Bender, Güneş Ragsdale, Stephen W. Drennan, Catherine L Massachusetts Institute of Technology. Department of Biology Massachusetts Institute of Technology. Department of Chemistry Ando, Nozomi Kung, Yan Drennan, Catherine L. In the Wood−Ljungdahl carbon fixation pathway, protein−protein interactions between methyltransferase (MeTr) and corrinoid iron−sulfur protein (CFeSP) are required for the transfer of a methyl group. While crystal structures have been determined for MeTr and CFeSP both free and in complex, solution structures have not been established. Here, we examine the transient interactions between MeTr and CFeSP in solution using anaerobic small-angle Xray scattering (SAXS) and present a global analysis approach for the deconvolution of heterogeneous mixtures formed by weakly interacting proteins. We further support this SAXS analysis with complementary results obtained by anaerobic isothermal titration calorimetry. Our results indicate that solution conditions affect the cooperativity with which CFeSP binds to MeTr, resulting in two distinct CFeSP/MeTr complexes with differing oligomeric compositions, both of which are active. One assembly resembles the CFeSP/MeTr complex observed crystallographically with 2:1 protein stoichiometry, while the other best fits a 1:1 CFeSP/MeTr arrangement. These results demonstrate the value of SAXS in uncovering the rich solution behavior of transient protein interactions visualized by crystallography. National Institutes of Health (U.S.) (NIH grant K99GM100008) National Institutes of Health (U.S.) (NIH grant F32GM090486) National Institutes of Health (U.S.) (NIH grant GM39451) National Institutes of Health (U.S.) (NIH grant GM69857) Howard Hughes Medical Institute (Investigator) 2013-11-07T16:31:50Z 2013-11-07T16:31:50Z 2012-10 2012-06 Article http://purl.org/eprint/type/JournalArticle 0002-7863 1520-5126 http://hdl.handle.net/1721.1/82017 Ando, Nozomi, Yan Kung, Mehmet Can, Gunes Bender, Stephen W. Ragsdale, and Catherine L. Drennan. “Transient B12-Dependent Methyltransferase Complexes Revealed by Small-Angle X-ray Scattering.” Journal of the American Chemical Society 134, no. 43 (October 31, 2012): 17945-17954. © 2012 American Chemical Society. https://orcid.org/0000-0001-5486-2755 en_US http://dx.doi.org/10.1021/ja3055782 Journal of the American Chemical Society Article is made available in accordance with the publisher's policy and may be subject to US copyright law. Please refer to the publisher's site for terms of use. application/pdf American Chemical Society ACS Author Choice |
| spellingShingle | Ando, Nozomi Kung, Yan Can, Mehmet Bender, Güneş Ragsdale, Stephen W. Drennan, Catherine L Transient B12-Dependent Methyltransferase Complexes Revealed by Small-Angle X-ray Scattering |
| title | Transient B12-Dependent Methyltransferase Complexes Revealed by Small-Angle X-ray Scattering |
| title_full | Transient B12-Dependent Methyltransferase Complexes Revealed by Small-Angle X-ray Scattering |
| title_fullStr | Transient B12-Dependent Methyltransferase Complexes Revealed by Small-Angle X-ray Scattering |
| title_full_unstemmed | Transient B12-Dependent Methyltransferase Complexes Revealed by Small-Angle X-ray Scattering |
| title_short | Transient B12-Dependent Methyltransferase Complexes Revealed by Small-Angle X-ray Scattering |
| title_sort | transient b12 dependent methyltransferase complexes revealed by small angle x ray scattering |
| url | http://hdl.handle.net/1721.1/82017 https://orcid.org/0000-0001-5486-2755 |
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