Interactions outside the proteinase-binding loop contribute significantly to the inhibition of activated coagulation factor XII by its canonical inhibitor from corn.

Autor:	Korneeva VA; From the Laboratory of the Molecular Mechanisms of Hemostasis, Center for Theoretical Problems of Physicochemical Pharmacology of Russian Academy of Sciences, Moscow 119991, Russia., Trubetskov MM; From the Laboratory of the Molecular Mechanisms of Hemostasis, Center for Theoretical Problems of Physicochemical Pharmacology of Russian Academy of Sciences, Moscow 119991, Russia, the Department of Physics, Moscow State University, Moscow 119992, Russia., Korshunova AV; the Research Department, HemaCore LLC, Moscow 125319, Russia., Lushchekina SV; the Laboratory of Computer Modeling of Biomolecular Systems and Nanomaterials, Emanuel Institute of Biochemical Physics of Russian Academy of Sciences, Moscow 119334, Russia., Kolyadko VN; From the Laboratory of the Molecular Mechanisms of Hemostasis, Center for Theoretical Problems of Physicochemical Pharmacology of Russian Academy of Sciences, Moscow 119991, Russia., Sergienko OV; the Laboratory of Molecular Diagnostics and Genetic Engineering, Institute of Agricultural Biotechnology of Russian Academy of Agricultural Sciences, Moscow 127550, Russia., Lunin VG; the Laboratory of Molecular Diagnostics and Genetic Engineering, Institute of Agricultural Biotechnology of Russian Academy of Agricultural Sciences, Moscow 127550, Russia, the Laboratory of Biologically Active Nanostructures, Gamaleya Institute of Epidemiology and Microbiology of Russian Federation Ministry of Health and Social Development, Moscow 123098, Russia., Panteleev MA; From the Laboratory of the Molecular Mechanisms of Hemostasis, Center for Theoretical Problems of Physicochemical Pharmacology of Russian Academy of Sciences, Moscow 119991, Russia, the Department of Physics, Moscow State University, Moscow 119992, Russia, the Research Department, HemaCore LLC, Moscow 125319, Russia, the Research Division, Scientific Clinical Centre of Pediatric Hematology, Oncology, and Immunology Named after Dmitry Rogachev of Ministry of Health of Russian Federation, Moscow 117997, Russia, and the Department of Translational and Regenerative Medicine, Moscow Institute of Physics and Technology, Dolgoprudny, Moscow Region, 141700, Russia mapanteleev@yandex.ru., Ataullakhanov FI; From the Laboratory of the Molecular Mechanisms of Hemostasis, Center for Theoretical Problems of Physicochemical Pharmacology of Russian Academy of Sciences, Moscow 119991, Russia, the Department of Physics, Moscow State University, Moscow 119992, Russia, the Research Department, HemaCore LLC, Moscow 125319, Russia, the Research Division, Scientific Clinical Centre of Pediatric Hematology, Oncology, and Immunology Named after Dmitry Rogachev of Ministry of Health of Russian Federation, Moscow 117997, Russia, and the Department of Translational and Regenerative Medicine, Moscow Institute of Physics and Technology, Dolgoprudny, Moscow Region, 141700, Russia.
Jazyk:	angličtina
Zdroj:	The Journal of biological chemistry [J Biol Chem] 2014 May 16; Vol. 289 (20), pp. 14109-20. Date of Electronic Publication: 2014 Apr 04.
DOI:	10.1074/jbc.M114.553735
Abstrakt:	Activated factor XII (FXIIa) is selectively inhibited by corn Hageman factor inhibitor (CHFI) among other plasma proteases. CHFI is considered a canonical serine protease inhibitor that interacts with FXIIa through its protease-binding loop. Here we examined whether the protease-binding loop alone is sufficient for the selective inhibition of serine proteases or whether other regions of a canonical inhibitor are involved. Six CHFI mutants lacking different N- and C-terminal portions were generated. CHFI-234, which lacks the first and fifth disulfide bonds and 11 and 19 amino acid residues at the N and C termini, respectively, exhibited no significant changes in FXIIa inhibition (Ki = 3.2 ± 0.4 nm). CHFI-123, which lacks 34 amino acid residues at the C terminus and the fourth and fifth disulfide bridges, inhibited FXIIa with a Ki of 116 ± 16 nm. To exclude interactions outside the FXIIa active site, a synthetic cyclic peptide was tested. The peptide contained residues 20-45 (Protein Data Bank code 1BEA), and a C29D substitution was included to avoid unwanted disulfide bond formation between unpaired cysteines. Surprisingly, the isolated protease-binding loop failed to inhibit FXIIa but retained partial inhibition of trypsin (Ki = 11.7 ± 1.2 μm) and activated factor XI (Ki = 94 ± 11 μm). Full-length CHFI inhibited trypsin with a Ki of 1.3 ± 0.2 nm and activated factor XI with a Ki of 5.4 ± 0.2 μm. Our results suggest that the protease-binding loop is not sufficient for the interaction between FXIIa and CHFI; other regions of the inhibitor also contribute to specific inhibition. (© 2014 by The American Society for Biochemistry and Molecular Biology, Inc.)
Databáze:	MEDLINE
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