Vitamin K-dependent carboxylation. A synthetic peptide based upon the gamma-carboxylation recognition site sequence of the prothrombin propeptide is an active substrate for the carboxylase in vitro.

Ulrich, Magda M. W.; Furie, B; Jacobs, MA; Vermeer, Cees

doi:10.1016/s0021-9258(19)81574-0

Cited by 91 publications

(31 citation statements)

References 41 publications

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“…9 It has been established that MK-4 is the main form of vitamin K in the brain. 9 The best known function of vitamin K 2 is to act as an essential cofactor for γ-glutamyl carboxylase, which catalyzes the posttranscriptional transformation of glutamate residues in γ-glutamate in some proteins, 10 enabling these proteins to better binding calcium ions. These modified proteins are involved in many physiological processes such as blood coagulation, bone homeostasis, and inhibition of vascular calcification, 11 but many other functions have been recently suggested for vitamins K. For example, much less studied is the role of both phylloquinone and menaquinone as antioxidant agents acting in cell membranes since, given their hydrophobic nature, they will be found in these structures preventing lipid peroxidation.…”

Section: ■ Introductionmentioning

confidence: 99%

Interaction of Vitamin K₁ and Vitamin K₂ with Dimyristoylphosphatidylcholine and Their Location in the Membrane

et al. 2020

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Vitamin K 1 and vitamin K 2 play very important biological roles as members of chains of electron transport as antioxidants in membranes and as cofactors for the posttranslational modification of proteins that participate in a number of physiological functions such as coagulation. The interaction of these vitamins with dimyristoylphosphatidylcholine (DMPC) model membranes has been studied by using a biophysical approach. It was observed by using differential scanning calorimetry that both vitamins have a very limited miscibility with DMPC and they form domains rich in the vitamins at high concentrations. Experiments using X-ray diffraction also showed the formation of different phases as a consequence of the inclusion of either vitamin K at temperatures below the phase transition. However, in the fluid state, a homogeneous phase was detected, and a decrease in the thickness of the membrane was accompanied by an increase in the water layer thickness. 2 H NMR spectroscopy showed that both vitamins K induced a decrease in the onset of the phase transition, which was bigger for vitamin K 1 , and both vitamins decreased the order of the membrane as seen through the first moment (M 1 ). 1 H NOESY MAS-NMR showed that protons located at the rings or at the beginning of the lateral chain of both vitamins K interacted with a clear preference with protons located in the polar part of DMPC. On the other hand, protons located on the lateral chain have a nearer proximity with the methyl end of the myristoyl chains of DMPC. In agreement with the 2 H NMR, ATR-FTIR (attenuated total reflectance Fourier transform infrared spectroscopy) indicated that both vitamins decreased the order parameters of DMPC. It was additionally deduced that the lateral chains of both vitamins were oriented almost in parallel to the myristoyl chains of the phospholipid.

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Section: ■ Introductionmentioning

confidence: 99%

Interaction of Vitamin K₁ and Vitamin K₂ with Dimyristoylphosphatidylcholine and Their Location in the Membrane

et al. 2020

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“…Other than MGP, which has an internal recognition site for carboxylase, all Gla-proteins characterized so far are synthesized as preproproteins [11]. The presequence is required for translocation across the endoplasmic reticulum membrane, whereas the prosequence functions as a high-affinity recognition site for carboxylase [12,13]. After carboxylation, the protein is transported through the cell to the trans-Golgi network.…”

Section: Introductionmentioning

confidence: 99%

“…With descarboxy-osteocalcin in purified propeptide-free recombinant carboxylase, the K m was 1.8 µM. Furthermore, osteocalcin was an inhibitor of descarboxy-osteocalcin carboxylation with a K i of Abbreviations used : bEELOMe, tripeptide t-butoxycarbonyl-Glu-Glu-Leu-OMe ; CHAPS,dimethylammonio]-2-hydroxy-1propanesulphonic acid ; d-OC, descarboxy-osteocalcin ; d-OC [13][14][15][16][17][18][19][20][21][22][23][24][25] and d-OC [22][23][24][25][26][27][28][29][30][31] , synthetic descarboxy-osteocalcin residues 13-25 and 22-31 respectively ;…”

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confidence: 99%

Osteocalcin binds tightly to the γ-glutamylcarboxylase at a site distinct from that of the other known vitamin K-dependent proteins

HOUBEN

Dayun

et al. 1999

Biochem. J.

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Vitamin K-dependent proteins contain a propeptide that is required for recognition by the enzyme gamma-glutamylcarboxylase. Substrates used in vitro for carboxylation studies lacking a prosequence are characterized by Km values in the millimolar range, whereas the Km for peptides containing a prosequence is three or four orders of magnitude smaller. Here we report that descarboxy-osteocalcin is an exception in this respect. With descarboxy-osteocalcin in purified propeptide-free recombinant carboxylase, the Km was 1.8 microM. Furthermore, osteocalcin was an inhibitor of descarboxy-osteocalcin carboxylation with a Ki of 76 microM. In contrast with the other vitamin K-dependent proteins, free propeptides do not inhibit descarboxy-osteocalcin carboxylation. Moreover, propeptide-containing substrates were inhibited neither by osteocalcin nor by its propeptide. From our studies we conclude that descarboxy-osteocalcin must have an internal recognition sequence that binds to gamma-glutamylcarboxylase at a site different from the propeptide-recognition site.

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“…In addition to the Glu-peptide-binding site, there is also a propeptide-binding site on carboxylase. The propeptide region (residues -18 to -1; Figure 2) of the precursor forms of the vitamin K dependent proteins confers 1000-fold tighter binding to Glu substrates when incorporated into the same synthetic peptide (Ulrich et al, 1988;Hubbard et al, 1989a). These propeptide residues are highly conserved (Pan & Price, 1985), and mutational analysis indicated that residues -18, -17, -16, -15, and -10 comprise the "7-carboxylation recognition site" (7-CRS) which directs the downstream and distant 7-carboxylation of the glutamates within the Gla domain (Ulrich et al, 1988;Huber et al, 1990).…”

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confidence: 99%

N-Bromoacetyl-peptide substrate affinity labeling of vitamin K dependent carboxylase

Kuliopulos¹,

Cieurzo²,

Furie³

et al. 1992

Biochemistry

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Vitamin K dependent carboxylase (carboxylase) is a membrane-associated endoplasmic reticular enzyme that catalyzes the conversion of certain glutamate residues of essential blood coagulation proteins to gamma-carboxyglutamyl (Gla) residues. A series of N-bromoacetyl-peptide substrate affinity labels based on the Gla domain of these blood-clotting proteins was synthesized, and the substrate and inactivator kinetic parameters were assessed. The most promising of these affinity peptides, N-bromoacetyl-FLEELY, was both substrate for carboxylase and an irreversible time-dependent inactivator of the enzyme, inactivating 80% of carboxylase under pseudo-first-order conditions. Addition of saturating amounts of a competing peptide substrate completely abolished the inhibitory properties of N-bromoacetyl-FLEELY, consistent with inactivation occurring at the active site. The partition ratio of inactivation/carboxylation was 1/30. The 94-kDa carboxylase was purified to 15-50% purity by a modification of a recent protocol [Wu, S.-M., Morris, D. P., & Stafford, D. W. (1991) Proc. Natl. Acad. Sci. U.S.A. 88, 2236-2240] and covalently labeled with N-bromoacetyl-FLEEL[125I]Y. On silver-stained 10% sodium dodecyl sulfate-polyacrylamide gels, the predominant radiolabeled band was the 94,000 molecular weight species. This result independently validates that the 94-kDa protein is a carboxylase.

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Vitamin K-dependent carboxylation. A synthetic peptide based upon the gamma-carboxylation recognition site sequence of the prothrombin propeptide is an active substrate for the carboxylase in vitro.

Cited by 91 publications

References 41 publications

Interaction of Vitamin K₁ and Vitamin K₂ with Dimyristoylphosphatidylcholine and Their Location in the Membrane

Interaction of Vitamin K₁ and Vitamin K₂ with Dimyristoylphosphatidylcholine and Their Location in the Membrane

Osteocalcin binds tightly to the γ-glutamylcarboxylase at a site distinct from that of the other known vitamin K-dependent proteins

N-Bromoacetyl-peptide substrate affinity labeling of vitamin K dependent carboxylase

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Vitamin K-dependent carboxylation. A synthetic peptide based upon the gamma-carboxylation recognition site sequence of the prothrombin propeptide is an active substrate for the carboxylase in vitro.

Cited by 91 publications

References 41 publications

Interaction of Vitamin K1 and Vitamin K2 with Dimyristoylphosphatidylcholine and Their Location in the Membrane

Interaction of Vitamin K1 and Vitamin K2 with Dimyristoylphosphatidylcholine and Their Location in the Membrane

Osteocalcin binds tightly to the γ-glutamylcarboxylase at a site distinct from that of the other known vitamin K-dependent proteins

N-Bromoacetyl-peptide substrate affinity labeling of vitamin K dependent carboxylase

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Interaction of Vitamin K₁ and Vitamin K₂ with Dimyristoylphosphatidylcholine and Their Location in the Membrane

Interaction of Vitamin K₁ and Vitamin K₂ with Dimyristoylphosphatidylcholine and Their Location in the Membrane