Pull-Down of Metalloproteins in Their Native States by Using Desthiobiotin-Based Probes |
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Authors: | Chinh Ngo Radhika Mehta Kanchan Aggarwal Audrey G. Fikes Dr. Ines C. Santos Dr. Sylvester M. Greer Prof. Dr. Emily L. Que |
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Affiliation: | Department of Chemistry, University of Texas at Austin, 105 E 24th St. Stop A5300, Austin, TX, 78712 USA |
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Abstract: | One-third of all proteins are estimated to require metals for structural stability and/or catalytic activity. Desthiobiotin probes containing metal binding groups can be used to capture metalloproteins with exposed active-site metals under mild conditions so as to prevent changes in metallation state. The proof-of-concept was demonstrated with carbonic anhydrase (CA), an open active site, Zn2+-containing protein. CA was targeted by using sulfonamide derivatives. Linkers of various lengths and structures were screened to determine the optimal structure for capture of the native protein. The optimized probes could selectively pull down CA from red blood cell lysate and other protein mixtures. Pull-down of differently metallated CAs was also investigated. |
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Keywords: | carbonic anhydrase desthiobiotin metalloproteins proteomics sulfonamide |
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