Substrate Recognition by Vitamin K-Dependent Carboxylase

SummaryDecarboxylated osteocalcins were prepared and purified from bovine, chicken, human and monkey bones and assayed for their ability to serve as a substrate for vitamin K-dependent carboxylase from bovine liver. Substantial differences were observed, especially between bovine and monkey d-osteocalcin. Since these substrates differ only in their amino acid residues 3 and 4, it seems that these residues play a role in the recognition of a substrate by hepatic carboxylase.

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Prediction of amino acid residues participated in substrate recognition by cytochrome P450 subfamilies with broad substrate specificity

Journal of Molecular Recognition ◽

10.1002/jmr.2251 ◽

2013 ◽

Vol 26 (2) ◽

pp. 86-91 ◽

Cited By ~ 5

Author(s):

Maria S. Zharkova ◽

Boris N. Sobolev ◽

Nina Yu. Oparina ◽

Alexander V. Veselovsky ◽

Alexander I. Archakov

Keyword(s):

Cytochrome P450 ◽

Amino Acid ◽

Substrate Specificity ◽

Substrate Recognition ◽

Amino Acid Residues ◽

Broad Substrate Specificity

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In vivo investigation of the substrate recognition capability and activity affecting amino acid residues of glycosyltransferase FscMI in the biosynthesis of candicidin

Molecular BioSystems ◽

10.1039/c2mb25464f ◽

2013 ◽

Vol 9 (3) ◽

pp. 422 ◽

Cited By ~ 5

Author(s):

Xuan Lei ◽

Lingxin Kong ◽

Chen Zhang ◽

Qian Liu ◽

Fen Yao ◽

...

Keyword(s):

Amino Acid ◽

Substrate Recognition ◽

Amino Acid Residues ◽

Recognition Capability

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Identification of amino acid residues involved in substrate recognition by the catalytic subunit of bovine cyclic AMP dependent protein kinase: peptide-based affinity labels

Biochemistry ◽

10.1021/bi00410a025 ◽

1988 ◽

Vol 27 (10) ◽

pp. 3691-3696 ◽

Cited By ~ 15

Author(s):

Shahriar Mobashery ◽

E. T. Kaiser

Keyword(s):

Amino Acid ◽

Protein Kinase ◽

Cyclic Amp ◽

Substrate Recognition ◽

Catalytic Subunit ◽

Amino Acid Residues ◽

Dependent Protein Kinase ◽

Affinity Labels ◽

Dependent Protein

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Contribution to Substrate Recognition of Two Aromatic Amino Acid Residues in Putative Transmembrane Segment 10 of the Yeast Sugar Transporters Gal2 and Hxt2

Journal of Biological Chemistry ◽

10.1074/jbc.273.44.29106 ◽

1998 ◽

Vol 273 (44) ◽

pp. 29106-29112 ◽

Cited By ~ 13

Author(s):

Michihiro Kasahara ◽

Mari Maeda

Keyword(s):

Amino Acid ◽

Aromatic Amino Acid ◽

Substrate Recognition ◽

Amino Acid Residues ◽

Transmembrane Segment ◽

Sugar Transporters

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Amino acid residues Arg659, Arg660, and Tyr661 in the spacer domain of ADAMTS13 are critical for cleavage of von Willebrand factor

Blood ◽

10.1182/blood-2009-07-235101 ◽

2010 ◽

Vol 115 (11) ◽

pp. 2300-2310 ◽

Cited By ~ 54

Author(s):

Sheng-Yu Jin ◽

Christopher G. Skipwith ◽

X. Long Zheng

Keyword(s):

Shear Stress ◽

Amino Acid ◽

Von Willebrand Factor ◽

Fluid Shear Stress ◽

Substrate Recognition ◽

Amino Acid Residues ◽

Fluid Shear ◽

Thrombocytopenic Purpura ◽

Von Willebrand ◽

Willebrand Factor

AbstractPrevious studies have shown that ADAMTS13 spacer domain is required for cleavage of von Willebrand factor (VWF). However, the exact amino acid residues within this domain critical for substrate recognition are not known. Epitope mapping of anti-ADAMTS13 immunoglobulin G from patients with thrombotic thrombocytopenic purpura and sequence alignment of the ADAMTS13 spacer domains of human, mouse, and zebrafish with these of human and murine ADAMTS1, a closely related member of ADAMTS family, have provided hints to investigate the role of the amino acid residues between Arg659 and Glu664 of the ADAMTS13 spacer domain in substrate recognition. A deletion of all these 6 amino acid residues (ie, Arg659-Glu664) from the ADAMTS13 spacer domain resulted in dramatically reduced proteolytic activity toward VWF73 peptides, guanidine-HCl denatured VWF, and native VWF under fluid shear stress, as well as ultralarge VWF on endothelial cells. Site-directed mutagenesis, kinetic analyses, and peptide inhibition assays have further identified a role for amino acid residues Arg659, Arg660, and Tyr661 in proteolytic cleavage of various substrates under static and fluid shear stress conditions. These findings may provide novel insight into the structural-function relationship of ADAMTS13 and help us to understand pathogenesis of thrombotic thrombocytopenic purpura and other arterial thromboses associated with compromised VWF proteolysis.

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cDNA cloning and tissue-specific expression of the phosphatidylcholine transfer protein gene

Biochemical Journal ◽

10.1042/bj3160049 ◽

1996 ◽

Vol 316 (1) ◽

pp. 49-55 ◽

Cited By ~ 19

Author(s):

Teunis B. H. GEIJTENBEEK ◽

Alexander J. SMITH ◽

Piet BORST ◽

Karel W. A. WIRTZ

Keyword(s):

Amino Acid ◽

Embryonic Development ◽

Amino Acid Sequence ◽

Bovine Liver ◽

Amino Acid Residues ◽

Adult Liver ◽

Bile Formation ◽

Rnase Protection ◽

Transfer Protein ◽

Phosphatidylcholine Transfer Protein

We have isolated a cDNA containing the complete coding sequence of bovine liver phosphatidylcholine transfer protein (PC-TP). The deduced amino acid sequence consists of 213 amino acid residues and is, except for a lysine instead of an arginine at position 167, identical to the sequence determined by Edman degradation [Akeroyd, Moonen, Westerman, Puyk and Wirtz (1981) Eur. J. Biochem. 114, 385–391]. A cDNA encoding amino acid residues 41–214 of mouse lung PC-TP was also isolated. The predicted amino acid sequence was 90% similar (81% identical) to the corresponding sequence of bovine liver PC-TP, demonstrating that PC-TP is conserved among mammalian species. By Southern blot analysis, evidence was obtained for the presence of a single bovine PC-TP-encoding gene. The expression of the PC-TP gene was determined during mouse embryonic development and in adult mouse tissues using an RNase protection assay. PC-TP RNA was present in embryos at all stages of development as early as the embryonic stem cell, suggesting a role for PC-TP in cell growth and differentiation. Towards the end of embryonic development, just before term, high levels of PC-TP RNA were found in the liver. This level was even higher 7 days post-term. In addition to adult liver, high levels of PC-TP RNA were also found in kidney and testis. The prominent presence of PC-TP in developing and adult liver is compatible with its proposed role in bile formation.

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The Carboxylation Of Bovine Vitamin K-Dependent Proteins By Bovine Carboxylase

10.1055/s-0038-1651977 ◽

1981 ◽

Author(s):

C Vermeer ◽

B A M Soute ◽

H C Hemker ◽

M de Metz

Keyword(s):

Amino Acid ◽

Vitamin K ◽

Specific Adsorption ◽

The Other ◽

Amino Acid Residues ◽

Factor X ◽

Good Substrate ◽

Enzyme Systems ◽

Endogenous Substrate ◽

Low Efficiency

Bovine vitamin K-dependent carboxylase was prepared, both from normal cows and from warfarin-treated ones. The two carboxylating enzyme systems were similar except for the fact that carboxylase from warfarin-treated cows contained a 100-fold higher amount of endogenous substrate. About 65% of this substrate consisted of one-chain factor X-precursor (s) and 25% of prothrombin-precursor(s). Solubilized carboxylase could be purified more than 100-fold by immuno- specific adsorption onto immobilized antifactor X antibodies. The purified enzyme contained 40% (w/w) phospholipid (mainly phosphatidylcholine), which was absolutely required for the carboxylating activity.Carboxylase from non-anticoagulated cows was used for studying exogenous substrates. Bovine descarboxyprothrombin, descarboxyfragment-1 and the pentapeptide Phe-Leu-Glu- Glu-Leu were carboxylated with a low efficiency (Km, between 0.4 and 11 mM). On the other hand a peptide, identical to the amino acid residues 13-29 in descarboxyprothrombin, had a Km of 0.001 mM. It seems probable therefore that the latter peptide contains all information required for a good substrate.

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