Axial iron ligation and protein encapsulation of the heme cofactor have been investigated as effectors of the reduction potential (Edegrees') of cytochrome c through direct electrochemistry experiments. Our approach was that of partitioning the Edegrees' changes resulting from binding of imidazole, 2-methyl-imidazole, ammonia, and azide to both cytochrome c and microperoxidase-11 (MP11), into the enthalpic and entropic contributions. N-Acetylmethionine binding to MP11 was also investigated. These ligands replace Met80 and a water molecule axially coordinated to the heme iron in cytochrome c and MP11, respectively. This factorization was achieved through variable temperature Edegrees' measurements. In this way, we have found that (1) the decrease in Edegrees' of cytochrome c due to Met80 substitution by a nitrogen-donor ligand is almost totally enthalpic in origin, as a result of the stronger electron donor properties of the exogenous ligand which selectively stabilize the ferric state; (ii) on the contrary, the binding of the same ligands and N-acetymethionine to MP11 results in an enthalpic stabilization of the reduced state, whereas the entropic effect invariably decreases Edegrees' (the former effect prevails for the methionine ligand and the latter for the nitrogenous ligands). A comparison of the reduction thermodynamics of cytochrome c and the MP11 adducts offers insight on the effect of changing axial heme ligation and heme insertion into the folded polypeptide chain. Principally, we have found that the overall Edegrees' increase of approximately 400 mV, comparing MP11 and native cytochrome c, consists of two opposite enthalpic and entropic terms of approximately +680 and -280 mV, respectively. The enthalpic term includes contributions from both axial methionine binding (+300 mV) and protein encapsulation of the heme (+380 mV), whereas the entropic term is almost entirely manifest at the stage of axial ligand binding. Both terms are dominated by the effects of water exclusion from the heme environment.

Control of cytochrome c redox potential: Axial ligation and protein environment effects / Battistuzzi, Gianantonio; Borsari, Marco; Ja, Cowan; Ranieri, Antonio; Sola, Marco. - In: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY. - ISSN 0002-7863. - STAMPA. - 124:19(2002), pp. 5315-5324. [10.1021/ja017479v]

Control of cytochrome c redox potential: Axial ligation and protein environment effects

BATTISTUZZI, Gianantonio;BORSARI, Marco;RANIERI, Antonio;SOLA, Marco
2002

Abstract

Axial iron ligation and protein encapsulation of the heme cofactor have been investigated as effectors of the reduction potential (Edegrees') of cytochrome c through direct electrochemistry experiments. Our approach was that of partitioning the Edegrees' changes resulting from binding of imidazole, 2-methyl-imidazole, ammonia, and azide to both cytochrome c and microperoxidase-11 (MP11), into the enthalpic and entropic contributions. N-Acetylmethionine binding to MP11 was also investigated. These ligands replace Met80 and a water molecule axially coordinated to the heme iron in cytochrome c and MP11, respectively. This factorization was achieved through variable temperature Edegrees' measurements. In this way, we have found that (1) the decrease in Edegrees' of cytochrome c due to Met80 substitution by a nitrogen-donor ligand is almost totally enthalpic in origin, as a result of the stronger electron donor properties of the exogenous ligand which selectively stabilize the ferric state; (ii) on the contrary, the binding of the same ligands and N-acetymethionine to MP11 results in an enthalpic stabilization of the reduced state, whereas the entropic effect invariably decreases Edegrees' (the former effect prevails for the methionine ligand and the latter for the nitrogenous ligands). A comparison of the reduction thermodynamics of cytochrome c and the MP11 adducts offers insight on the effect of changing axial heme ligation and heme insertion into the folded polypeptide chain. Principally, we have found that the overall Edegrees' increase of approximately 400 mV, comparing MP11 and native cytochrome c, consists of two opposite enthalpic and entropic terms of approximately +680 and -280 mV, respectively. The enthalpic term includes contributions from both axial methionine binding (+300 mV) and protein encapsulation of the heme (+380 mV), whereas the entropic term is almost entirely manifest at the stage of axial ligand binding. Both terms are dominated by the effects of water exclusion from the heme environment.
2002
124
19
5315
5324
Control of cytochrome c redox potential: Axial ligation and protein environment effects / Battistuzzi, Gianantonio; Borsari, Marco; Ja, Cowan; Ranieri, Antonio; Sola, Marco. - In: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY. - ISSN 0002-7863. - STAMPA. - 124:19(2002), pp. 5315-5324. [10.1021/ja017479v]
Battistuzzi, Gianantonio; Borsari, Marco; Ja, Cowan; Ranieri, Antonio; Sola, Marco
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11380/306493
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