X-ray structure of bovine heart cytochrome c at high ionic strength
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Bovine heart cytochrome c (bCyt c) is an extensively studied hemoprotein of only 104 residues. Due to the existence of isoforms generated by non-enzymatic deaminidation, crystallization of bCyt c is difficult and involves extensive purification and the use of microseeding or the presence of an electric field. Taking advantage of the capacity of cytochrome c (cyt c) to bind anions on its protein surface, the commercially available bCyt c was crystallized without extra purifications, using ammonium sulfate as precipitant and nitrate ions as additives. The structure of the ferric bCyt c in a new crystal form is described and compared with that previously solved at low ionic strength and with those of human and horse cyt c. The overall structure of bCyt c is conserved, while the side chains of several residues that play a role in the interactions of cyt c with its partners have different rotamers in the two structures. The effect of the presence of nitrate ions on the structure of the protein is then evaluated and compared with that observed in the case of ferrous and ferric horse heart cyt c.
KeywordsHemoprotein crystallization X-ray structure Cytochrome c Hot spots Protein–protein recognition
The author acknowledges students for his lab for technical assistance and members of ESRF staff for their help with data collection and processing.
Compliance with ethical standard
Conflict of interest
The author declares that he has no conflict of interest.
- De Rocco D, Cerqua C, Np Goffrini, Russo G, Pastore A, Meloni F, Nicchia E, Moraes CT, Pecci A, Salviati L et al (2014) Mutations of cytochrome c identified in patients with thrombocytopenia THC4 affect boith apoptosis and cellular bioenergetics. Biochim Biophys Acta Mol Basis Dis 1842:269–274CrossRefGoogle Scholar
- Josephs TM, Liptak MD, Hughes G, Lo A, Smith RM, Wilbanks SM, Bren KL, Ledgerwood EC (2013) Conformational change and human cytochrome c function: mutation of residue 41 modulates caspase activation and destabilizes Met-80 coordination. J Biol Inorg Chem 18(3):289–297CrossRefPubMedPubMedCentralGoogle Scholar
- Monari S, Battistuzzi G, Borsari M, Millo D, Gooijer C, van der Zwan G, Ranieri A (2008) Sola M (2008) Thermodynamic and kinetic aspects of the electron transfer reaction of bovine cytochrome c immobilized on 4-mercaptopyridine and 11-mercapto-1-undecanoic acid films. J Appl Electrochem 38:885–891CrossRefGoogle Scholar
- Moreno-Beltrán B, Guerra-Castellano A, Díaz-Quintana A, Del Conte R, García-Mauriño SM, Díaz-Moreno S, González-Arzola K, Santos-Ocaña C, Velázquez-Campoy A, De la Rosa MA, Turano P, Díaz-Moreno I (2017) Structural basis of mitochondrial dysfunction in response to cytochrome c phosphorylation at tyrosine 48. Proc Natl Acad Sci USA 114(15):E3041–E3050CrossRefPubMedPubMedCentralGoogle Scholar
- Rajagopal BS, Edzuma AN, Hough MA, Blundell KLIM, Kagan VE, Kapralov AA, Fraser LA, Butt JN, Silkstone GG, Wilson MT, Svistunenko DA, Worrall JAR (2013) The hydrogen peroxide induced radical behaviour in human cytochrome c phospholipid complexes: implications for the enhanced pro-apoptotic activity of the G41S mutant. Biochem J 456:441–452CrossRefPubMedGoogle Scholar
- Schrodinger LLC (2015) The PyMOL molecular graphics system. Version, 1 (8). www.pymol.org
- Scott RA, Mauk GA (eds) (1996) Cytochrome c: a multidisciplinary approach. University Science Books, SausalitoGoogle Scholar
- Winn MD, Ballard CC, Cowtan KD, Dodson EJ, Emsley P, Evans PR, Keegan RM, Krissinel EB, Leslie AGW, McCoy A, McNicholas SJ, Murshudov GN, Pannu NS, Potterton EA, Powell HR, Read RJ, Vagin A, Wilson KS (2011) Overview of the CCP4 suite and current developments. Acta Crystallogr Sect D Biol Crystallogr 67:235–242CrossRefGoogle Scholar