title: The phosphodiesterase activity of the HmsP EAL domain is required for negative regulation of biofilm formation in Yersinia pestis.
abstract: In Yersinia pestis, biofilm formation is stimulated by HmsT, a GGDEF-domain containing protein that synthesizes cyclic-di-GMP (c-di-GMP), and inhibited by HmsP, an EAL-domain protein. Only the EAL-domain portion of HmsP is required to inhibit biofilm formation. The EAL domain of HmsP was purified as a 6XHis-tag fusion protein and demonstrated to have phosphodiesterase activity using bis(p-nitrophenyl) phosphate (bis-pNPP) as a substrate. This enzymatic activity was strictly manganese dependent. A critical residue (E506) of HmsP within the EAL domain, that is required for inhibition of biofilm formation, is also essential for this phosphodiesterase activity. While the proposed function of EAL-domain proteins is to linearize c-di-GMP, this is a direct demonstration of the required phosphodiesterase activity of a purified EAL-domain protein.
authors.affiliation: Department of Microbiology, Immunology and Molecular Genetics, University of Kentucky, Lexington, KY 40536-0298, USA.
authors.forename: Alexander G
authors.initials: AG
authors.lastname: Bobrov
authors.forename: Olga
authors.initials: O
authors.lastname: Kirillina
authors.forename: Robert D
authors.initials: RD
authors.lastname: Perry
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D018441:Biofilms / Q000254:growth & development*; D003067:Coenzymes / Q000494:pharmacology; D015536:Down-Regulation; D008345:Manganese / Q000494:pharmacology; D009596:Nitrophenols / Q000378:metabolism; D010727:Phosphoric Diester Hydrolases / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D017434:Protein Structure, Tertiary; D015010:Yersinia pestis / Q000201:enzymology* / Q000235:genetics / Q000502:physiology
chemical_list: D001426:Bacterial Proteins; D003067:Coenzymes; D009596:Nitrophenols; D008345:Manganese; C002887:bis(4-nitrophenyl)phosphate; D010727:Phosphoric Diester Hydrolases