Isolation and N‐terminal amino acid sequence of protein Z, A γ‐carboxyglutamic acid containing protein from bovine plasma

Petersen, Torben E.; Thøgersen, Hans C.; Sottrup‐Jensen, Lars; Magnusson, Staffan; Jörnvall, Hans

doi:10.1016/0014-5793(80)81133-1

Cited by 30 publications

(13 citation statements)

References 21 publications

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“…We found, as noted by others for bovine Protein Z (4,6), that the use of an agarose-(as opposed to a dextran-) based ionexchange support was required for optimal separation of the Protein Z. The bars in Fig.…”

Section: Functional Assayssupporting

confidence: 69%

“…The amino acid composition of human Protein Z is shown in Table I, as is that previously published for bovine Protein Z (6). The NH2-terminal amino acid sequence is shown in Table II; for comparison, those of human Factors II, VII, IX, X, Protein C, Protein S, and bovine Protein Z (25)(26)(27)(28)(29)(30)(31) are also shown.…”

Section: Functional Assaysmentioning

confidence: 90%

“…Recently Prowse and Esnouf described another vitamin K-dependent protein in bovine plasma and they named it Protein Z (4). Initially this protein was thought to represent a form of Factor X (single chain), but it was later identified as a discrete gamma carboxyglutamic acid-containing protein (5,6). The physiological function of bovine Protein Z remains unknown.…”

Section: Introductionmentioning

confidence: 99%

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Human Protein Z.

Broze¹,

Miletich²

1984

J. Clin. Invest.

140

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A s bstract. Protein Z was purified from human plasma by a four-step procedure which included barium citrate adsorption, ammonium sulfate fractionation, DEAE-Sepharose chromatography, and blue agarose chromatography with a yield of 20%. It is a 62,000 mol wt protein with an extinction coefficient of 12.0. The concentration of Protein Z in pooled, citrated plasma is 2.2 ug/ml and its half-life in patients starting warfarin anticoagulation therapy is estimated to be <2.5 d. The NH2-terminal sequence is Ala-Gly-Ser-Tyr-Leu-Leu-(Gla)-(Gla)-Leu-Phe-(Gla)-Gly-Asn-Leu. Neither Protein Z nor its cleavage products, which were obtained by treatment ofProtein Z with thrombin or plasmin, incorporated [3H]diisopropyl fluorophosphate. The physiological function of Protein Z remains unknown.

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Section: Functional Assayssupporting

confidence: 69%

Section: Functional Assaysmentioning

confidence: 90%

Section: Introductionmentioning

confidence: 99%

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Human Protein Z.

Broze¹,

Miletich²

1984

J. Clin. Invest.

140

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“…In 1977, Prowse and Esnouf (4) identified an additional vitamin K-dependent protein circulating in bovine plasma and named it protein Z (PZ). Initially thought to represent a single-chain form of bovine factor X, bovine PZ was later identified as a discrete Glacontaining protein (5,6). The human counterpart of bovine PZ was isolated in 1984 (7).…”

mentioning

confidence: 99%

Isolation of a protein Z-dependent plasma protease inhibitor

Han

Fiehler

Broze

1998

Proc. Natl. Acad. Sci. U.S.A.

140

185

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Human protein Z (PZ) is a 62,000-M r , vitamin K-dependent plasma protein whose structure is similar to coagulation factors VII, IX, X, protein C, and protein S, but whose function is not known. The procoagulant activity of factor Xa in a one-stage plasma coagulation assay is reduced when factor Xa is first incubated with PZ. This apparent inhibitory effect is time dependent, requires the presence of calcium ions and procoagulant phospholipids (rabbit brain cephalin), and appears predominantly related to the incubation period of PZ with cephalin. In serum the initial rate of inhibition of factor Xa with calcium ions and cephalin also is enhanced in the presence PZ. A PZ-dependent protease inhibitor (ZPI) has been isolated from plasma. ZPI is a 72,000-M r single-chain protein with an N-terminal amino acid sequence of LAPSPQSPEXXA (X ‫؍‬ indeterminate) and an estimated concentration in citrate-treated plasma of 1.0-1.6 g͞ml. In systems using purified components, the factor Xa inhibition produced by ZPI is rapid (>95% within 1 min by coagulation assay) and requires the presence of PZ, calcium ions, and cephalin. The inhibitory process appears to involve the formation of a factor Xa-PZ-ZPI complex at the phospholipid surface.Vitamin K is required for the posttranslational formation of ␥-carboxyglutamic acid (Gla), which is present in a number of plasma proteins that are involved in coagulation: prothrombin, factors VII, IX, and X, protein C, and protein S (1, 2). Gla-mediated calcium ion binding in these proteins is necessary for their association with phospholipid surfaces and is critical for their hemostatic function (3). In 1977, Prowse and Esnouf (4) identified an additional vitamin K-dependent protein circulating in bovine plasma and named it protein Z (PZ). Initially thought to represent a single-chain form of bovine factor X, bovine PZ was later identified as a discrete Glacontaining protein (5, 6). The human counterpart of bovine PZ was isolated in 1984 (7).Human PZ is a 62,000-M r glycoprotein that has a plasma half-life of Ϸ2.5 days (8). Plasma PZ levels in blood donors span a broad range with a mean concentration of 2.9 Ϯ 1.0 g͞ml in EDTA-anticoagulated samples (corresponding to Ϸ2.6 g͞ml in citrated plasma) (8). The N-terminal half of PZ is very homologous to those of factors VII, IX, and X, and contains a Gla-domain, two epidermal growth factor-like domains, and a region that connects to a homologue of the catalytic domains present in the serine protease zymogens. In the C-terminal domain of PZ, however, the region around the typical ''activation'' site is absent and the His and Ser residues of the catalytic triad are lacking (the Asp residue is conserved) (9, 10).McDonald et al. (11) recently reported that the kinetics of the binding of human and bovine PZ to phosphatidylcholine͞ phosphatidylserine (75%͞25%) vesicles is different from that of the other vitamin K-dependent coagulation factors. The k assn (10) and k dssn (s Ϫ1 ) rate constants are 1.95 and 0.0063 for bovine PZ and 3.36 and 0.057 ...

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“…Purification of bovine Factor X was carried out by adsorption on barium citrate followed by ion-exchange chromatography as described for the isolation of bovine protein Z (Petersen et al, 1980 The freeze-dried fractions were digested with trypsin [1:20 (w/w) in 0.1 M-ammonium acetate buffer, pH 5.2, overnight at 37°C] or pepsin [1:20 (w/w) in 2% (v/v) acetic acid, overnight at 37°C]. Subdigests were carried out on isolated peptides with the following enzymes and conditions: trypsin (as above), thermolysin (in 10 mmCaCl2/100 mM-pyridinium acetate buffer, pH 6.5, 3-16 h at 37°C) or staphylococcal proteinase (in 100 mMammonium acetate buffer, pH 4.0, overnight at 37°C).…”

Section: Materials and Methods Materialsmentioning

confidence: 99%

Disulphide bridges of bovine factor X

Højrup

Magnusson

1987

Biochemical Journal

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Evidence is presented for the disulphide bridges in bovine Factor X. The protein was degraded by chemical and enzymic means, and all 12 disulphide bridges were isolated in separate peptides except for bridges nos. 6/7 in the light chain. All the disulphide bridges were found to be in positions corresponding to those found in other homologous domains. This report is the first verification of an epidermal-growth-factor-homologous domain having the same disulphide-bonding pattern as that found in mouse epidermal growth factor.

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Isolation and N‐terminal amino acid sequence of protein Z, A γ‐carboxyglutamic acid containing protein from bovine plasma

Cited by 30 publications

References 21 publications

Human Protein Z.

Human Protein Z.

Isolation of a protein Z-dependent plasma protease inhibitor

Disulphide bridges of bovine factor X

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