PDBsum entry 5jw1

Oxidoreductase

PDB id: 5jw1

Contents

Protein chains

552 a.a.

Ligands

NAG-NAG ×2

COH ×2

CEL ×2

BOG

NAG ×4

Waters ×15

References listed in PDB file

Key reference

• Title: Fatty acid binding to the allosteric subunit of cyclooxygenase-2 relieves a tonic inhibition of the catalytic subunit.
• Authors: L.Dong, C.Yuan, B.J.Orlando, M.G.Malkowski, W.L.Smith.
• Ref.: J Biol Chem, 2016, 291, 25641-25655. [DOI no: 10.1074/jbc.M116.757310]
• PubMed id: 27756840

Abstract

Prostaglandin endoperoxide H synthase-2 (PGHS-2), also called cyclooxygenase-2 (COX-2), converts arachidonic acid to PGH₂ PGHS-2 is a conformational heterodimer composed of allosteric (E_allo) and catalytic (E_cat) subunits. Fatty acids (FAs) bind to Arg-120 of E_allo increasing to different degrees, depending on the FA, the V_max of its E_cat partner. We report here that movement of helical residues 120-122 and loop residues 123-129 of E_allo underlies the allosteric effects of FAs and allosteric COX-2 inhibitors, including naproxen and flurbiprofen. An S121P substitution in both PGHS-2 monomers yields a variant (S121P/S121P PGHS-2) that has 1.7-1.8 times the V_max of native PGHS-2 and is relatively insensitive to activation by FAs or inhibition by allosteric inhibitors. The S121P substitution in E_allo is primarily responsible for these effects. In X-ray crystal structures, the Cα atoms of helical residues 119-122 of S121P/S121P PGHS-2 are displaced from their normal positions. Additionally, the S121P/S121P PGHS-2 variants in which Pro-127 and Ser-541 are replaced by cysteines spontaneously forms Cys-127 to Cys-541 cross-links between monomers. This is unlike the corresponding native PGHS-2 variant and suggests that S121P substitutions also unhinge the loop involving residues 123-129. We conclude the following: (a) the region involving residues 120-129 of unoccupied E_allo tonically inhibits E_cat; (b) binding of an activating FA (e.g. arachidonic, palmitic, or oleic acid) to E_allo or an S121P substitution in E_allo repositions this region to increase E_cat activity; and (c) allosteric COX inhibitors act by preventing FA binding to E_allo and additionally by relocating E_allo residues to inhibit E_cat.