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Publication Abstract from PubMed

CAMP factor is a unique alpha-helical bacterial toxin that is known for its co-hemolytic activity in combination with staphylococcal sphingomyelinase. It was first discovered in the human pathogen Streptococcus agalactiae (also known as group B streptococcus), but homologous genes have been found in many other Gram-positive pathogens. In this study, the efforts that led to the determination of the first structure of a CAMP-family toxin are reported. Initially, it was possible to produce crystals of the native protein which diffracted to near 2.45 A resolution. However, a series of technical obstacles were encountered on the way to structure determination. Over a period of more than five years, many methods, including selenomethionine labeling, mutations, crystallization chaperones and heavy-atom soaking, were attempted, but these attempts resulted in limited progress. The structure was finally solved using a combination of iodine soaking and molecular replacement using the crystallization chaperone maltose-binding protein (MBP) as a search model. Analysis of native and MBP-tagged CAMP-factor structures identified a conserved interaction interface in the C-terminal domain (CTD). The positively charged surface may be critical for binding to acidic ligands. Furthermore, mutations on the interaction interface at the CTD completely abolished its co-hemolytic activities. This study provides novel insights into the mechanism of the membrane-permeabilizing activity of CAMP factor.

Structure determination of the CAMP factor of Streptococcus agalactiae with the aid of an MBP tag and insights into membrane-surface attachment.,Li Y, Zeng W, Li Y, Fan W, Ma H, Fan X, Jiang J, Brefo-Mensah E, Zhang Y, Yang M, Dong Z, Palmer M, Jin T Acta Crystallogr D Struct Biol. 2019 Aug 1;75(Pt 8):772-781. doi:, 10.1107/S205979831901057X. Epub 2019 Jul 31. PMID:31373576^[1]

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.