Suppr超能文献

维生素 K 依赖性蛋白前体原肽、维生素 K 对苯二酚和谷氨酸底物结合对 γ-谷氨酰羧化酶结构和功能的影响。

Effect of vitamin K-dependent protein precursor propeptide, vitamin K hydroquinone, and glutamate substrate binding on the structure and function of {gamma}-glutamyl carboxylase.

机构信息

Department of Biology, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.

出版信息

J Biol Chem. 2010 Oct 8;285(41):31502-8. doi: 10.1074/jbc.M110.143297. Epub 2010 Aug 17.

Abstract

The γ-glutamyl carboxylase utilizes four substrates to catalyze carboxylation of certain glutamic acid residues in vitamin K-dependent proteins. How the enzyme brings the substrates together to promote catalysis is an important question in understanding the structure and function of this enzyme. The propeptide is the primary binding site of the vitamin K-dependent proteins to carboxylase. It is also an effector of carboxylase activity. We tested the hypothesis that binding of substrates causes changes to the carboxylase and in turn to the substrate-enzyme interactions. In addition we investigated how the sequences of the propeptides affected the substrate-enzyme interaction. To study these questions we employed fluorescently labeled propeptides to measure affinity for the carboxylase. We also measured the ability of several propeptides to increase carboxylase catalytic activity. Finally we determined the effect of substrates: vitamin K hydroquinone, the pentapeptide FLEEL, and NaHCO(3), on the stability of the propeptide-carboxylase complexes. We found a wide variation in the propeptide affinities for carboxylase. In contrast, the propeptides tested had similar effects on carboxylase catalytic activity. FLEEL and vitamin K hydroquinone both stabilized the propeptide-carboxylase complex. The two together had a greater effect than either alone. We conclude that the effect of propeptide and substrates on carboxylase controls the order of substrate binding in such a way as to ensure efficient, specific carboxylation.

摘要

γ-谷氨酰羧化酶利用四种底物,催化维生素 K 依赖性蛋白中某些谷氨酸残基的羧化作用。酶如何将底物聚集在一起以促进催化是理解该酶结构和功能的一个重要问题。前肽是维生素 K 依赖性蛋白与羧化酶的主要结合位点。它也是羧化酶活性的效应物。我们检验了这样一个假设,即底物的结合会引起羧化酶的变化,并进一步引起底物-酶相互作用的变化。此外,我们还研究了前肽序列如何影响底物-酶相互作用。为了研究这些问题,我们使用荧光标记的前肽来测量与羧化酶的亲和力。我们还测量了几种前肽增加羧化酶催化活性的能力。最后,我们确定了底物:维生素 K 氢醌、五肽 FLEEL 和 NaHCO(3)对前肽-羧化酶复合物稳定性的影响。我们发现前肽与羧化酶的亲和力有很大差异。相比之下,测试的前肽对羧化酶的催化活性有相似的影响。FLEEL 和维生素 K 氢醌都能稳定前肽-羧化酶复合物。两者一起的效果比单独使用任何一种都要好。我们的结论是,前肽和底物对羧化酶的影响控制着底物结合的顺序,以确保有效的、特异性的羧化作用。

相似文献

引用本文的文献

1
Structural insights into the vitamin K-dependent γ-carboxylation of osteocalcin.
Cell Res. 2025 Sep 2. doi: 10.1038/s41422-025-01161-0.
2
Sustained high expression of human FVII following AAV8-mediated gene delivery in mice.
Mol Ther Methods Clin Dev. 2025 Jun 25;33(3):101523. doi: 10.1016/j.omtm.2025.101523. eCollection 2025 Sep 11.
3
The Molecular Basis of FIX Deficiency in Hemophilia B.
Int J Mol Sci. 2022 Mar 2;23(5):2762. doi: 10.3390/ijms23052762.
5
Insights into vitamin K-dependent carboxylation: home field advantage.
Haematologica. 2020 Aug;105(8):1996-1998. doi: 10.3324/haematol.2020.253690.
6
Vitamin K-dependent carboxylation of coagulation factors: insights from a cell-based functional study.
Haematologica. 2020 Aug;105(8):2164-2173. doi: 10.3324/haematol.2019.229047. Epub 2019 Oct 17.
7
The WAGR syndrome gene PRRG4 is a functional homologue of the commissureless axon guidance gene.
PLoS Genet. 2017 Aug 31;13(8):e1006865. doi: 10.1371/journal.pgen.1006865. eCollection 2017 Aug.
8
Functional Study of the Vitamin K Cycle Enzymes in Live Cells.
Methods Enzymol. 2017;584:349-394. doi: 10.1016/bs.mie.2016.10.015. Epub 2016 Nov 22.
9
Characterization of vitamin K-dependent carboxylase mutations that cause bleeding and nonbleeding disorders.
Blood. 2016 Apr 14;127(15):1847-55. doi: 10.1182/blood-2015-10-677633. Epub 2016 Jan 12.
10
Structural and functional insights into enzymes of the vitamin K cycle.
J Thromb Haemost. 2016 Feb;14(2):236-47. doi: 10.1111/jth.13217. Epub 2016 Jan 29.

本文引用的文献

2
Quantum chemical study of the mechanism of action of vitamin K carboxylase (VKC). IV. Intermediates and transition states.
J Phys Chem A. 2007 Aug 9;111(31):7257-61. doi: 10.1021/jp068564y. Epub 2007 May 16.
3
A quantum chemical study of the mechanism of action of Vitamin K carboxylase (VKC) III. Intermediates and transition states.
J Mol Graph Model. 2007 Sep;26(2):409-14. doi: 10.1016/j.jmgm.2006.10.006. Epub 2006 Nov 6.
8
A novel fluorescence assay to study propeptide interaction with gamma-glutamyl carboxylase.
Biochemistry. 2001 Oct 2;40(39):11723-33. doi: 10.1021/bi010332w.

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验