Pull‐Down of Metalloproteins in Their Native States by Using Desthiobiotin‐Based Probes. (25th February 2019)

• Record Type: Journal Article
• Authors: Ngo, Chinh; Mehta, Radhika; Aggarwal, Kanchan; Fikes, Audrey G.; Santos, Ines C.; Greer, Sylvester M.; Que, Emily L.
• Is Part Of: Chembiochem. Volume 20:Number 8(2019)
• Page Start: 1003
• Page End: 1007
• Publication Date: 2019-02-25
• Subjects: carbonic anhydrase -- desthiobiotin -- metalloproteins -- proteomics -- sulfonamide; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.201800613
• Languages: English
• ISSNs: 1439-4227

Abstract

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, Zn 2+ ‐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. Abstract : Avoiding the destruct button : One‐third of all proteins are estimated to require metals for structural stability and/or catalytic activity. Desthiobiotin probes with metal‐binding sulfonamide groups enable the pull‐down of the metalloprotein carbonic anhydrase under mild conditions without destroying the protein metallation state.