4Z74

Crystal structure of inorganic pyrophosphatase from Mycobacterium tuberculosis in complex with inorganic pyrophosphate

Summary for 4Z74

• Entry DOI: 10.2210/pdb4z74/pdb
• Related: 4Z70 4Z71 4Z72 4Z73
• Descriptor: Inorganic pyrophosphatase, PYROPHOSPHATE 2-, CALCIUM ION, ... (4 entities in total)
• Functional Keywords: pyrophosphatase, phosphatase, hydrolase, inorganic pyrophosphate
• Biological source: Mycobacterium tuberculosis (strain ATCC 25618 / H37Rv)
• Cellular location: Cytoplasm : P9WI55
• Total number of polymer chains: 12
• Total formula weight: 237333.34

Authors

Pratt, A.C.,Biswas, T.,Tsodikov, O.V. (deposition date: 2015-04-06, release date: 2015-08-26, Last modification date: 2024-03-06)

Primary citation

Pratt, A.C.,Dewage, S.W.,Pang, A.H.,Biswas, T.,Barnard-Britson, S.,Cisneros, G.A.,Tsodikov, O.V.
Structural and computational dissection of the catalytic mechanism of the inorganic pyrophosphatase from Mycobacterium tuberculosis.
J.Struct.Biol., 192:76-87, 2015

PubMed Abstract: Family I inorganic pyrophosphatases (PPiases) are ubiquitous enzymes that are critical for phosphate metabolism in all domains of life. The detailed catalytic mechanism of these enzymes, including the identity of the general base, is not fully understood. We determined a series of crystal structures of the PPiase from Mycobacterium tuberculosis (Mtb PPiase) bound to catalytic metals, inorganic pyrophosphate (PPi; the reaction substrate) and to one or two inorganic phosphate ions (Pi; the reaction product), ranging in resolution from 1.85 to 3.30Å. These structures represent a set of major kinetic intermediates in the catalytic turnover pathway for this enzyme and suggest an order of association and dissociation of the divalent metals, the substrate and the two products during the catalytic turnover. The active site of Mtb PPiase exhibits significant structural differences from the well characterized Escherichia coli PPiase in the vicinity of the bound PPi substrate. Prompted by these differences, quantum mechanics/molecular mechanics (QM/MM) analysis yielded an atomic description of the hydrolysis step for Mtb PPiase and, unexpectedly, indicated that Asp89, rather than Asp54 that was proposed for E. coli PPiase, can abstract a proton from a water molecule to activate it for a nucleophilic attack on the PPi substrate. Mutagenesis studies of the key Asp residues of Mtb PPiase supported this mechanism. This combination of structural and computational analyses clarifies our understanding of the mechanism of family I PPiases and has potential utility for rational development of drugs targeting this enzyme.

PubMed: 26296329
DOI: 10.1016/j.jsb.2015.08.010

Experimental method

X-RAY DIFFRACTION (2.55 Å)