title: Structure and cooperativity of a T-state mutant of histidine decarboxylase from Lactobacillus 30a.
abstract: Histidine decarboxylase (HDC) from Lactobacillus 30a converts histidine to histamine, a process that enables the bacteria to maintain the optimum pH range for cell growth. HDC is regulated by pH; it is active at low pH and inactive at neutral to alkaline pH. The X-ray structure of HDC at pH 8 revealed that a helix was disordered, resulting in the disruption of the substrate-binding site. The HDC trimer has also been shown to exhibit cooperative kinetics at neutral pH, that is, histidine can trigger a T-state to R-state transition. The D53,54N mutant of HDC has an elevated Km, even at low pH, indicating that the enzyme assumes the low activity T-state. We have solved the structures of the D53,54N mutant at low pH, with and without the substrate analog histidine methyl ester (HME) bound. Structural analysis shows that the apo-D53,54N mutant is in the inactive or T-state and that binding of the substrate analog induces the enzyme to adopt the active or R-state. A mechanism for the cooperative transition is proposed.
authors.affiliation: Institute of Cellular and Molecular Biology, Department of Chemistry and Biochemistry, University of Texas, Austin, Texas 78712, USA.
authors.forename: Scott
authors.initials: S
authors.lastname: Worley
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Schelp
authors.forename: Arthur F
authors.initials: AF
authors.lastname: Monzingo
authors.forename: Stephen
authors.initials: S
authors.lastname: Ernst
authors.forename: Jon D
authors.initials: JD
authors.lastname: Robertus
mesh_terms: D019943:Amino Acid Substitution; D006639:Histidine / Q000031:analogs & derivatives* / Q000737:chemistry; D006640:Histidine Decarboxylase / Q000737:chemistry / Q000235:genetics*; D007700:Kinetics; D007778:Lactobacillus / Q000737:chemistry / Q000235:genetics*; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D011487:Protein Conformation
chemical_list: D006639:Histidine; C053861:histidine methyl ester; D006640:Histidine Decarboxylase