Mechanistic Scrutiny Identifies a Kinetic Role for Cytochrome b5 Regulation of Human Cytochrome P450c17 (CYP17A1, P450 17A1)

Mechanistic Scrutiny Identifies a Kinetic Role for Cytochrome b5 Regulation of Human Cytochrome P450c17 (CYP17A1, P450 17A1) Full article

Общее

Language: Английский, Genre: Full article,
Status: Published, Source type: Original

Journal

PLoS One
ISSN: 1932-6203

Output data

Year: 2015, Volume: 10, Number: 11, Article number : e0141252, Pages count : 19 DOI: 10.1371/journal.pone.0141252

Authors

Simonov Alexandr N.

¹ , Holien Jessica K. ² , Yeung Joyee Chun In ¹ , Nguyen Ann D. ³ , Corbin C.Jo ³ , Zheng Jie ⁴ , Kuznetsov Vladimir L.

⁵ , Auchus Richard J. ⁶ , Conley Alan J. ³ , Bond Alan M. ¹ , Parker Michael W. ^2,7 , Rodgers Raymond J. ⁸ , Martin Lisandra L. ¹

Affiliations

1	School of Chemistry, Monash University, Clayton, Victoria, Australia
2	ACRF Rational Drug Discovery Centre, St. Vincent's Institute of Medical Research, Fitzroy, Victoria, Australia
3	School of Veterinary Medicine, University of California Davis, Davis, California, United States of America
4	Department of Physiology and Membrane Biology, School of Medicine, University of California Davis, Davis, California, United States of America
5	Boreskov Institute of Catalysis, Prospekt Lavrentieva 5, Novosibirsk, Russia
6	Division of Metabolism, Endocrinology and Diabetes, Department of Internal Medicine, University of Michigan, Ann Arbor, Michigan, United States of America
7	Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, Victoria, Australia
8	Discipline of Obstetrics and Gynaecology, School of Paediatrics and Reproductive Health, Robinson Research Institute, University of Adelaide, Adelaide, South Australia, Australia

Funding (3)

1	National Institutes of Health	R01GM086596
2	Australian Research Council
3	National Health and Medical Research Council

Cytochrome P450c17 (P450 17A1, CYP17A1) is a critical enzyme in the synthesis of androgens and is now a target enzyme for the treatment of prostate cancer. Cytochrome P450c17 can exhibit either one or two physiological enzymatic activities differentially regulated by cytochrome b5. How this is achieved remains unknown. Here, comprehensive in silico, in vivo and in vitro analyses were undertaken. Fluorescence Resonance Energy Transfer analysis showed close interactions within living cells between cytochrome P450c17 and cytochrome b5. In silico modeling identified the sites of interaction and confirmed that E48 and E49 residues in cytochrome b5 are essential for activity. Quartz crystal microbalance studies identified specific protein-protein interactions in a lipid membrane. Voltammetric analysis revealed that the wild type cytochrome b5, but not a mutated, E48G/E49G cyt b5, altered the kinetics of electron transfer between the electrode and the P450c17. We conclude that cytochrome b5 can influence the electronic conductivity of cytochrome P450c17 via allosteric, protein-protein interactions.

Simonov A.N. , Holien J.K. , Yeung J.C.I. , Nguyen A.D. , Corbin C.J. , Zheng J. , Kuznetsov V.L. , Auchus R.J. , Conley A.J. , Bond A.M. , Parker M.W. , Rodgers R.J. , Martin L.L.
Mechanistic Scrutiny Identifies a Kinetic Role for Cytochrome b5 Regulation of Human Cytochrome P450c17 (CYP17A1, P450 17A1)

PLoS One. 2015. V.10. N11. e0141252 :1-19. DOI: 10.1371/journal.pone.0141252 publication_identifier_short.wos_identifier_type publication_identifier_short.scopus_identifier_type publication_identifier_short.rinz_identifier_type

Full text from publisher

Submitted:	Jul 27, 2015
Accepted:	Oct 6, 2015
Published print:	Nov 20, 2015
Published online:	Nov 20, 2015

publication_identifier.wos_identifier_type	WOS:000365251500003
publication_identifier.scopus_identifier_type	2-s2.0-84958025929
publication_identifier.rinz_identifier_type	26824785
publication_identifier.accession_number_identifier_type	2016:331
publication_identifier.chemical_accession_number_identifier_type	164:246940