Involvement of porin N,N-dicyclohexylcarbodiimide-reactive domain in hexokinase binding to the outer mitochondrial membrane
- PMID: 15756812
- DOI: 10.1007/s10930-004-0600-2
Involvement of porin N,N-dicyclohexylcarbodiimide-reactive domain in hexokinase binding to the outer mitochondrial membrane
Abstract
The proportion of hexokinase that is bound to the outer mitochondrial membrane is tissue specific and metabolically regulated. This study examined the role of the N,N-dicyclohexylcarbodiimide-binding domain of mitochondrial porin in binding to hexokinase 1. Selective proteolytic cleavage of porin protein was performed and peptides were assayed for their, effect on hexokinase I binding to isolated mitochondria. Specificity of DCCD-reactive domain binding to hexokinase I was demonstrated by competition of the peptides for porin binding sites on hexokinase as well as by blockage hexokinase binding by N,N-dicyclohexylcarbodiimide. One of the peptides, designated as 5 kDa (the smallest of the porin peptides, which contains a DCCD-reactive site), totally blocked binding of the enzyme to the mitochondrial membrane, and significantly enhanced the release of the mitochondrially bound enzyme. These experiments demonstrate that there exists a direct and specific interaction between the DCCD-reactive domain of VDAC and hexokinase I. The peptides were further characterized with respect to their effects on certain functional properties of hexokinase I. None had any detectable effect on catalytic properties, including inhibition by glucose 6-phosphate. To evaluate further the outer mitochondrial membrane's role in the hexokinase binding, insertion of VDAC was examined using isolated rat mitochondria. Preincubation of mitochondria with purified porin strongly increases hexokinase I binding to rat liver mitochondria. Collectively, the results imply that the high hexokinase-binding capability of porin-enriched mitochondria was due to a quantitative difference in binding sites.
Similar articles
-
Hexokinase receptor complex in hepatoma mitochondria: evidence from N,N'-dicyclohexylcarbodiimide-labeling studies for the involvement of the pore-forming protein VDAC.Biochemistry. 1986 Mar 11;25(5):1015-21. doi: 10.1021/bi00353a010. Biochemistry. 1986. PMID: 3008816
-
Hexokinase-binding properties of the mitochondrial VDAC protein: inhibition by DCCD and location of putative DCCD-binding sites.J Bioenerg Biomembr. 1989 Aug;21(4):461-70. doi: 10.1007/BF00762518. J Bioenerg Biomembr. 1989. PMID: 2478532 Review.
-
Porin proteins in mitochondria from rat pancreatic islet cells and white adipocytes: identification and regulation of hexokinase binding by the sulfonylurea glimepiride.Arch Biochem Biophys. 1994 Jan;308(1):8-23. doi: 10.1006/abbi.1994.1002. Arch Biochem Biophys. 1994. PMID: 8311478
-
Homophilic anchorage of brain-hexokinase to mitochondria-porins revealed by specific-peptide antibody cross recognition.Bull Cancer. 2004 Jun;91(6):E184-200. Bull Cancer. 2004. PMID: 15562563
-
[Cooperation of membrane proteins and cytosolic proteins in metabolic regulation--involvement of binding of hexokinase to mitochondria in regulation of glucose metabolism and association and complex formation between membrane proteins and cytosolic proteins in regulation of active oxygen production].Yakugaku Zasshi. 1999 Jan;119(1):16-34. doi: 10.1248/yakushi1947.119.1_16. Yakugaku Zasshi. 1999. PMID: 9922708 Review. Japanese.
Cited by
-
Hexokinase 2 is a determinant of neuroblastoma metastasis.Br J Cancer. 2016 Mar 29;114(7):759-66. doi: 10.1038/bjc.2016.26. Epub 2016 Mar 17. Br J Cancer. 2016. PMID: 26986252 Free PMC article.
-
VDAC regulation of mitochondrial calcium flux: From channel biophysics to disease.Cell Calcium. 2021 Mar;94:102356. doi: 10.1016/j.ceca.2021.102356. Epub 2021 Jan 23. Cell Calcium. 2021. PMID: 33529977 Free PMC article. Review.
-
VDAC Regulation: A Mitochondrial Target to Stop Cell Proliferation.Adv Cancer Res. 2018;138:41-69. doi: 10.1016/bs.acr.2018.02.002. Epub 2018 Mar 2. Adv Cancer Res. 2018. PMID: 29551129 Free PMC article. Review.
-
Targeting mitochondrial ion channels for cancer therapy.Redox Biol. 2021 Jun;42:101846. doi: 10.1016/j.redox.2020.101846. Epub 2020 Dec 24. Redox Biol. 2021. PMID: 33419703 Free PMC article. Review.
-
Regulation of hexokinase binding to VDAC.J Bioenerg Biomembr. 2008 Jun;40(3):171-82. doi: 10.1007/s10863-008-9148-8. J Bioenerg Biomembr. 2008. PMID: 18683036 Free PMC article. Review.
References
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources