2IUX

Human tACE mutant g1234

Summary for 2IUX

• Entry DOI: 10.2210/pdb2iux/pdb
• Related: 1O86 1O8A 1UZE 1UZF 2IUL
• Descriptor: ANGIOTENSIN-CONVERTING ENZYME, 2-acetamido-2-deoxy-beta-D-glucopyranose, ACETATE ION, ... (7 entities in total)
• Functional Keywords: glycosidase, polymorphism, glycoprotein, metal-binding, alternative splicing, metalloprotease, phosphorylation, ac, zinc, membrane, chloride, protease, hydrolase, type-i membrane-anchored protein, carboxypeptidase, peptidyl dipeptidase, transmembrane
• Biological source: HOMO SAPIENS (HUMAN)
• Total number of polymer chains: 1
• Total formula weight: 69096.12

Authors

Watermeyer, J.M.,Swell, B.T.,Natesh, R.,Corradi, H.R.,Acharya, K.R.,Sturrock, E.D. (deposition date: 2006-06-07, release date: 2006-10-25, Last modification date: 2020-07-29)

Primary citation

Watermeyer, J.M.,Sewell, B.T.,Schwager, S.L.,Natesh, R.,Corradi, H.R.,Acharya, K.R.,Sturrock, E.D.
Structure of Testis Ace Glycosylation Mutants and Evidence for Conserved Domain Movement.
Biochemistry, 45:12654-, 2006

PubMed Abstract: Human angiotensin-converting enzyme is an important drug target for which little structural information has been available until recent years. The slow progress in obtaining a crystal structure was due to the problem of surface glycosylation, a difficulty that has thus far been overcome by the use of a glucosidase-1 inhibitor in the tissue culture medium. However, the prohibitive cost of these inhibitors and incomplete glucosidase inhibition makes alternative routes to minimizing the N-glycan heterogeneity desirable. Here, glycosylation in the testis isoform (tACE) has been reduced by Asn-Gln point mutations at N-glycosylation sites, and the crystal structures of mutants having two and four intact sites have been solved to 2.0 A and 2.8 A, respectively. Both mutants show close structural identity with the wild-type. A hinge mechanism is proposed for substrate entry into the active cleft, based on homology to human ACE2 at the levels of sequence and flexibility. This is supported by normal-mode analysis that reveals intrinsic flexibility about the active site of tACE. Subdomain II, containing bound chloride and zinc ions, is found to have greater stability than subdomain I in the structures of three ACE homologues. Crystallizable glycosylation mutants open up new possibilities for cocrystallization studies to aid the design of novel ACE inhibitors.

PubMed: 17042482
DOI: 10.1021/BI061146Z

Experimental method

X-RAY DIFFRACTION (2.8 Å)