1KRC

CRYSTAL STRUCTURE OF KLEBSIELLA AEROGENES UREASE, ITS APOENZYME AND TWO ACTIVE SITE MUTANTS

Summary for 1KRC

• Entry DOI: 10.2210/pdb1krc/pdb
• Descriptor: UREASE, NICKEL (II) ION, CARBON DIOXIDE, ... (6 entities in total)
• Functional Keywords: active site mutant, nickel metalloenzyme, hydrolase (urea amido)
• Biological source: Klebsiella aerogenes; More
• Cellular location: Cytoplasm (By similarity): P18316 P18315 P18314
• Total number of polymer chains: 3
• Total formula weight: 83273.85

Authors

Jabri, E.,Karplus, P.A. (deposition date: 1995-06-20, release date: 1995-10-15, Last modification date: 2024-06-05)

Primary citation

Jabri, E.,Karplus, P.A.
Structures of the Klebsiella aerogenes urease apoenzyme and two active-site mutants.
Biochemistry, 35:10616-10626, 1996

PubMed Abstract: Urease from Klebsiella aerogenes [Jabri et al. (1995) Science 268, 998-1004] is an (alpha beta gamma)3 trimer with each alpha-subunit having an (alpha beta)8-barrel domain containing a binickel active center. Here we examine structure-function relations for urease in more detail through structural analysis of the urease apoenzyme at 2.3 A resolution and mutants of two key catalytic residues (H219A and H320A) at 2.5 A resolution. With the exception of the active site, in which a water molecule takes the place of the missing carbamate and nickel atoms, the structure of the apoenzyme is nearly identical to that of the holoenzyme, suggesting a high degree of preorganization which helps explain the tight binding of nickel. In the structure of H219A, the major change involves a conformational shift and ordering of the active site flap, but a small shift in the side chain of Asp alpha 221 could contribute to the lower activity of H219A. In the H320A structure, the catalytic water, primarily a Ni-2 ligand in the holoenzyme, shifts into a bridging position. This shift shows that the nickel ligation is rather sensitive to the environment and the change in ligation may contribute to the 10(5)-fold lower activity of H320A. In addition, these results show that urease is resilient to the loss of nickel ions and mutations. Analysis of the urease tertiary/quaternary structure suggests that the stability of this enzyme may be largely due to its burial of an unusually large fraction of its residues: 50% in the gamma-subunit, 30% in the beta-subunit, and 60% in the alpha-subunit.

PubMed: 8718850
DOI: 10.1021/bi960424z

Experimental method

X-RAY DIFFRACTION (2.5 Å)