2A2S

Crystal Structure of Human Glutathione Transferase in complex with S-nitrosoglutathione in the absence of reducing agent

Summary for 2A2S

• Entry DOI: 10.2210/pdb2a2s/pdb
• Related: 1ZGN 2A2R 5GSS
• Descriptor: Glutathione S-transferase P, 2-(N-MORPHOLINO)-ETHANESULFONIC ACID, 2-AMINO-5-[1-(CARBOXYLATOMETHYLCARBAMOYL)-2-NITROSOSULFANYL-ETHYL]AMINO-5-OXO-PENTANOATE, ... (6 entities in total)
• Functional Keywords: transferase, detoxification, nitric oxide carrier, s-nitrosoglutathione
• Biological source: Homo sapiens (human)
• Cellular location: Cytoplasm : P09211
• Total number of polymer chains: 2
• Total formula weight: 47914.71

Authors

Parker, L.J.,Morton, C.J.,Adams, J.J.,Parker, M.W. (deposition date: 2005-06-23, release date: 2006-06-06, Last modification date: 2023-10-25)

Primary citation

Tellez-Sanz, R.,Cesareo, E.,Nuccetelli, M.,Aguilera, A.M.,Baron, C.,Parker, L.J.,Adams, J.J.,Morton, C.J.,Lo Bello, M.,Parker, M.W.,Garcia-Fuentes, L.
Calorimetric and structural studies of the nitric oxide carrier S-nitrosoglutathione bound to human glutathione transferase P1-1
Protein Sci., 15:1093-1105, 2006

PubMed Abstract: The nitric oxide molecule (NO) is involved in many important physiological processes and seems to be stabilized by reduced thiol species, such as S-nitrosoglutathione (GSNO). GSNO binds strongly to glutathione transferases, a major superfamily of detoxifying enzymes. We have determined the crystal structure of GSNO bound to dimeric human glutathione transferase P1-1 (hGSTP1-1) at 1.4 A resolution. The GSNO ligand binds in the active site with the nitrosyl moiety involved in multiple interactions with the protein. Isothermal titration calorimetry and differential scanning calorimetry (DSC) have been used to characterize the interaction of GSNO with the enzyme. The binding of GSNO to wild-type hGSTP1-1 induces a negative cooperativity with a kinetic process concomitant to the binding process occurring at more physiological temperatures. GSNO inhibits wild-type enzyme competitively at lower temperatures but covalently at higher temperatures, presumably by S-nitrosylation of a sulfhydryl group. The C47S mutation removes the covalent modification potential of the enzyme by GSNO. These results are consistent with a model in which the flexible helix alpha2 of hGST P1-1 must move sufficiently to allow chemical modification of Cys47. In contrast to wild-type enzyme, the C47S mutation induces a positive cooperativity toward GSNO binding. The DSC results show that the thermal stability of the mutant is slightly higher than wild type, consistent with helix alpha2 forming new interactions with the other subunit. All these results suggest that Cys47 plays a key role in intersubunit cooperativity and that under certain pathological conditions S-nitrosylation of Cys47 by GSNO is a likely physiological scenario.

PubMed: 16597834
DOI: 10.1110/ps.052055206

Experimental method

X-RAY DIFFRACTION (1.7 Å)