2BN7

Mn substituted E. coli Aminopeptidase P in complex with product and Zn

Summary for 2BN7

• Entry DOI: 10.2210/pdb2bn7/pdb
• Related: 1A16 1JAW 1M35 1N51 1W2M 1W7V 1WBQ 1WL6 1WL9 1WLR 2BH3 2BHA 2BHB 2BHC 2BHD 2BWS 2BWT 2BWU 2BWV 2BWW 2BWX 2BWY
• Descriptor: XAA-PRO AMINOPEPTIDASE, PROLINE, LEUCINE, ... (8 entities in total)
• Functional Keywords: hydrolase, proline-specific peptidase, product complex, metalloenzyme, pita-bread fold, dinuclear hydrolase
• Biological source: ESCHERICHIA COLI
• Total number of polymer chains: 1
• Total formula weight: 50379.05

Authors

Graham, S.C.,Bond, C.S.,Freeman, H.C.,Guss, J.M. (deposition date: 2005-03-22, release date: 2005-09-29, Last modification date: 2023-12-13)

Primary citation

Graham, S.C.,Bond, C.S.,Freeman, H.C.,Guss, J.M.
Structural and Functional Implications of Metal Ion Selection in Aminopeptidase P, a Metalloprotease with a Dinuclear Metal Center.
Biochemistry, 44:13820-13836, 2005

PubMed Abstract: The effect of metal substitution on the activity and structure of the aminopeptidase P (APPro) from Escherichia coli has been investigated. Measurements of activity in the presence of Mn2+, Mg2+, Zn2+, Na+, and Ca2+ show that significant activity is seen only in the Mn-bound form of the enzyme. The addition of Zn2+ to [MnMn(APPro)] is strongly inhibitory. Crystal structures of [MnMn(APPro)], [MgMg(APPro)], [ZnZn(APPro)], [ZnMg(APPro)], [Ca_(APPro)], [Na_(APPro)], and [apo(APPro)] were determined. The structures of [Ca_(APPro)] and [Na_(APPro)] have a single metal atom at their active site. Surprisingly, when a tripeptide substrate (ValProLeu) was soaked into [Na_(APPro)] crystals in the presence of 200 mM Mg2+, the structure had substrate, but no metal, bound at the active site. The structure of apo APPro complexed with ValProLeu shows that the N-terminal amino group of a substrate can be bound at the active site by carboxylate side chains that normally bind the second metal atom, providing a model for substrate binding in a single-metal active enzyme. Structures of [MnMn(APPro)] and [ZnZn(APPro)] complexes of ProLeu, a product inhibitor, in the presence of excess Zn reveal a third metal-binding site, formed by two conserved His residues and the dipeptide inhibitor. A Zn atom bound at such a site would stabilize product binding and enhance inhibition.

PubMed: 16229471
DOI: 10.1021/BI0512849

Experimental method

X-RAY DIFFRACTION (2.4 Å)