Mass-spectrometric identification of interaction sites for cytochrome P450 2b4/nadph cytochrome P450 reductase

Ivanov A.V.1, Kopylov A.T.1, Zgoda V.G.1, Toropygin I.Yu.1, Khrjapova E.V.1, Ivanov Yu.D.1

1. Institute of Biomedical Chemistry, Russian Academy of Medical Sciences
Section: Experimental/Clinical Study
DOI: 10.18097/PBMC20105601040      PubMed Id: 21328910
Year: 2010  Volume: 56  Issue: 1  Pages: 40-54
We determined the interaction sites of the cytochrome P450's protein-partners: 2B4 (d-2B4) and NADPH-cytochrome P450 of reductase (d-Fp). While in operation, these proteins are forming the complexes. We used 4-4'-dithio(bisphenyl)azide linker for non-specific covalent coupling of d-2B4 complexes with d-Fp in Emulgen-913 - monomerized system. Covalently-linked peptides in this complex were identified with ESI-MS/MS. Several sites of these proteins' binding with each other were revealed. Based on them, a model of intermolecular protein interactions was created. The model includes 5 cross-linker-stabilized contact sites of d-2B4 with d-Fp involving the following peptides of d-2B4 and d-Fp: 1) d-2B4423-433 и d-Fp 102-109; 2) d-2B4324-336 и d-Fp570-585; 3) d-2B4327-336 и d-Fp452-464; 4) d-2B4 192-197 и d-Fp456-464; 5) d-2B4 134-139 и d-Fp406-425.Herein, in the latter two cases, the peptides of d-Fp are located in their inter-domain slit and stabilize protein-protein complex via nanoprobe cross-linker; therefore, the formation of d-2B4/d-Fp complexes in these sites may involve aminoacid residues d-Fp456-464 and d-Fp406-425 surrounding inter-domain slit.
Download PDF:  
Keywords: cytochrome P450 2B4, NADPH cytochrome P450 reductase, mass-spectrometry

Ivanov, A. V., Kopylov, A. T., Zgoda, V. G., Toropygin, I. Yu., Khrjapova, E. V., Ivanov, Yu. D. (2010). Mass-spectrometric identification of interaction sites for cytochrome P450 2b4/nadph cytochrome P450 reductase. Biomeditsinskaya Khimiya, 56(1), 40-54.
This paper is also available as the English translation: 10.1134/S1990750809040052
 2024 (vol 70)
 2023 (vol 69)
 2022 (vol 68)
 2021 (vol 67)
 2020 (vol 66)
 2019 (vol 65)
 2018 (vol 64)
 2017 (vol 63)
 2016 (vol 62)
 2015 (vol 61)
 2014 (vol 60)
 2013 (vol 59)
 2012 (vol 58)
 2011 (vol 57)
 2010 (vol 56)
 2009 (vol 55)
 2008 (vol 54)
 2007 (vol 53)
 2006 (vol 52)
 2005 (vol 51)
 2004 (vol 50)
 2003 (vol 49)