A Novel Dysfibrinogenemia with Abnormal γ-Chain(Fibrinogen Nagoya).

1979 ◽  
Author(s):  
Junki Takamatsu ◽  
Kanji Oqata ◽  
Tadashi Kamiya ◽  
Katsuo Koie ◽  
Takagi Takashi ◽  
...  
Keyword(s):  
Amino Acid ◽  
Inhibitory Effect ◽  
Chain Structure ◽  
Japanese Family ◽  
Electrophoretic Mobilities ◽  
Sds Page ◽  
History Of ◽  
The Difference ◽  
Fibrin Monomers ◽  
Major Defect

Six individuals in 3 generations of Japanese family had prolonged thrombin clotting time, but no history of hemorrhagic or thrombotic disease. Very low fibrinogen levels were obtained by thrombin clottable protein, while immunological procedures gave normal values of fibrinogen. The serum contained 40-80μg/ml of unclottable fibrinogen related antigens. The patients’ plasma had an inhibitory effect on the fibrin formation in normal plasma. Major defect of this fibrinogen was a delayed aggregation of fibrin monomers.On CM-chromatography (CM-52) of the S-carboxymethylated fibrinogen, three different γ-chains, named γx, γL and γR, were separated. They did not differ in their electrophoretic mobilities in SDS-PAGE, but were distinguishable in PAGE containing 8M urea. Moreover, the amino acid compositions and tryptic peptide mappings of each chain revealed a little difference from those of normal fibrinogen γ chains, suggesting the difference in amino acid substitution or oligosaccharide chain structure.Based on these findings, we designated this fibrinogen as fibrinogen Nagoya; its possible identity without other dysfibrinogenemia has not been excluded.

Purification and Characterization of Lupus Anticoagulant Like Protein from Agkistrodon Halys Brevicaudus Venom

1996 ◽  
Vol 76 (06) ◽  
pp. 0993-0997
Author(s):  
Zhao-Yan Li ◽  
Xiao-Wei Wu ◽  
Tie-Fu Yu ◽  
Eric C-Y Lian
Keyword(s):  
Amino Acid ◽  
Lupus Anticoagulant ◽  
Inhibitory Effect ◽  
Clotting Factor ◽  
Acid Oxidase ◽  
Crude Venom ◽  
Amino Acid Oxidase ◽  
Sds Page ◽  
The Individual ◽  
Reducing Condition

SummaryBy means of CM-Sephadex C-25, DEAE-Sephadex A-50, Sephadex G-200, and Sephadex G-75 chromatographies, a lupus anticoagulant like protein (LALP) from Agkistrodon halys brevicaudus was purified. On SDS-PAGE, the purified LALP had a molecular weight of 25,500 daltons under non-reducing condition and 15,000 daltons under reducing condition. The isoelectric point was pH 5.6. Its N terminal amino acid sequencing revealed a mixture of 2 sequences: DCP(P/S)(D/G)WSSYEGH(C/R)Q(Q/K). It was devoid of phospho-lipaseA, fibrino(geno)lytic, 5′-nucleotidase, L-amino acid oxidase, phosphomonoesterase, phosphodiesterase and thrombin-like activities, which were found in crude venom. In the presence of LALP, PT, aPTT, and dRVVT of human plasma were markedly prolonged and its effects were concentration-dependent but time-independent. The inhibitory effect of LALP on the plasma clotting time was enhanced by decreasing phospholipid concentration in TTI test. The individual clotting factor activity was not affected by LALP when higher dilutions of LALP-plasma mixture were used for assay. Russell’s viper venom time was shortened when high phospholipid confirmatory reagent was used. Therefore, the protein has lupus anticoagulant property.

Fibrinogen Alès: a homozygous case of dysfibrinogenemia (γ-Asp330 →Val) characterized by a defective fibrin polymerization site “a”

Blood ◽  
2000 ◽  
Vol 96 (10) ◽  
pp. 3473-3479 ◽  
Author(s):  
Karim Chabane Lounes ◽  
Claudine Soria ◽  
Shah Sultan Mirshahi ◽  
Pierre Desvignes ◽  
Massoud Mirshahi ◽  
...  
Keyword(s):  
Base Change ◽  
Molecular Defect ◽  
Fibrinopeptide A ◽  
Fibrin Polymerization ◽  
Prolonged Incubation ◽  
Fibrinogen Binding ◽  
Sds Page ◽  
History Of ◽  
Single Base Change ◽  
Fibrin Monomers

Abstract Congenital homozygous dysfibrinogenemia was diagnosed in a man with a history of 2 thrombotic strokes before age 30. His hemostatic profile was characterized by a dramatically prolonged plasma thrombin clotting time, and no clotting was observed with reptilase. Complete clotting of the abnormal fibrinogen occurred after a prolonged incubation of plasma with thrombin. The release of fibrinopeptides A and B by thrombin and of fibrinopeptide A by reptilase were both normal. Thrombin-induced fibrin polymerization was impaired, and no polymerization occurred with reptilase. The polymerization defect was characterized by a defective site “a,” resulting in an absence of interaction between sites A and a, indicated by the lack of fragment D1 (or fibrinogen) binding to normal fibrin monomers depleted in fibrinopeptide A only (Des-AA fm). By SDS-PAGE, the defect was detected on the γ-chain and in its fragment D1. The molecular defect determined by analysis of genomic DNA showed a single base change (A→T) in exon VIII of the γ-chain. The resulting change in the amino acid structure is γ 330 aspartic acid (GAT) → valine (GTT). It is concluded that the residue γ-Asp330 is essential for the normal functioning of the polymerization site a on the fibrinogen γ-chain.

scholarly journals Measurement report: Hydrolyzed amino acids in fine and coarse atmospheric aerosol in Nanchang, China: concentrations, compositions, sources and possible bacterial degradation state

2021 ◽  
Vol 21 (4) ◽  
pp. 2585-2600
Author(s):  
Ren-Guo Zhu ◽  
Hua-Yun Xiao ◽  
Li Luo ◽  
Hongwei Xiao ◽  
Zequn Wen ◽  
...  
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Biomass Burning ◽  
Fine Particles ◽  
Bacterial Degradation ◽  
Jiangxi Province ◽  
Coarse Particles ◽  
History Of ◽  
Composition Profiles ◽  
The Difference

Abstract. Amino acids (AAs) are relevant for nitrogen cycles, climate change and public health. Their size distribution may help to uncover the source, transformation and fate of protein in the atmosphere. This paper explores the use of compound-specific δ15N patterns of hydrolyzed amino acid (HAA), δ15N values of total hydrolyzed amino acid (δ15NTHAA), degradation index (DI) and the variance within trophic AAs (∑V) as markers to examine the sources and processing history of different sizes of particle in the atmosphere. Two weeks of daily aerosol samples from five sampling sites in the Nanchang area (Jiangxi Province, China) and samples of main emission sources of AAs in aerosols (biomass burning, soil and plants) were collected (Zhu et al., 2020). Here, we measured the concentrations and δ15N values of each HAA in two size-segregated aerosol particles (> 2.5 µm and PM2.5). Our results showed that the average concentrations of THAA in fine particles was nearly 6 times higher than that in coarse particles (p < 0.01) and composition profiles of fine and coarse particles were quite different from each other. The δ15N values of hydrolyzed glycine and THAA in both fine and coarse particles were typically in the range of those from biomass burning, soil and plant sources. Moreover, the average difference in the δ15NTHAA value between fine and coarse particles was smaller than 1.5 ‰. These results suggested that the sources of atmospheric HAAs for fine and coarse particles might be similar. Meanwhile, compared to fine particles, significantly lower DI values (p  <  0.05), “scattered” δ15N distribution in trophic AA and higher ∑V values (p < 0.05) were observed in coarse particles. But the difference in δ15N values of source AA (glycine, serine, phenylalanine and lysine) and THAA between coarse particles and fine particles was relatively small. It is likely that AAs in coarse particles have advanced bacterial degradation states compared to fine particles. Besides that, the significant increase in DI values and a decrease in ∑V values for coarse particles were observed on days on which precipitation fell (p  <  0.05). This implies that “fresh” AAs in coarse particles were likely released following the precipitation.

FIBRINOGENS KYOTO AND TOCHIGI, EACH WITH AN APPARENT ABNORMAL MOL. WT. γ CHAIN, ARE CHARACTERIZED BY REPLACEMENT OF γ ASN-308 BY LYS AND γ ARG-275 BY CYS, RESPECTIVELY

1987 ◽  
Author(s):  
N Yoshida ◽  
S Terukina ◽  
M Matsuda ◽  
M Moroi ◽  
M Okuma ◽  
...  
Keyword(s):  
Amino Acid ◽  
Sequence Analysis ◽  
Amino Acid Sequence ◽  
Cross Linking ◽  
Short Peptides ◽  
Amino Acid Sequence Analysis ◽  
Fibrin Monomer ◽  
Sds Page ◽  
Fibrin Monomers

Congenital inherited abnormal fibrinogens (Fbgs) Kyoto and Tochigi showed prolonged thrombin- and reptilase-time, normal release of fibrinopeptides A and B, normal cross linking ability and impaired polymerization of the fibrin monomer.Purified Fbg analyzed on SDS-PAGE under the reduced condition in the system of Laemmli contained 50 % of an apparent lower mol. wt. γ chain (γ Kyoto)(mol. wt.= 48,000 compared with 50,000 for the normal) in Fbg Kyoto and an apparent higher mol. wt. γ chain (γ Tochigi)(mol. wt.= 50,500) in Fbg Tochigi. Apparent mol. wt. differences were also detected in reduced and carboxymethyl ated Fbg, Fbg fragment D1, and D2, but not in D3. This suggested that the abnormality of γ chains in both Fbgs is in γ 303-356.Amino acid sequence analysis was performed for CNBr- or lysylendopeptidase-digested peptides of the γ chain or D1 peptides after fractionation on HPLC. In Fbg Kyoto, γ Asn-308 was substituted by Lys, and a deletion of short peptides corresponding to the mol. wt. difference of 2,000 could not be detected. In Fbg Tochigi, γ Arg-275 was substituted by Cys, and no abnormality of amino acid sequence was found in γ 303-356.These results suggest that some lesions or conformations containing γ 275 and γ 308 will directly or indirectly affect polymerization of fibrin monomers. Although the reason for apparent mol. wt. differences is not known yet, SDS-PAGE in the system of Laemmli will be useful for the analysis of abnormal Fbgs.Fbg Kyoto could not be separated into two or three populations and may contain hetero-dimer molecules, but Fbg Tochigi had unclottable Fbg with predominant γ Tochigi and may contain abnormal homo-dimer molecules and normal molecules.

Fibrinogen Alès: a homozygous case of dysfibrinogenemia (γ-Asp330 →Val) characterized by a defective fibrin polymerization site “a”

Blood ◽  
2000 ◽  
Vol 96 (10) ◽  
pp. 3473-3479
Author(s):  
Karim Chabane Lounes ◽  
Claudine Soria ◽  
Shah Sultan Mirshahi ◽  
Pierre Desvignes ◽  
Massoud Mirshahi ◽  
...  
Keyword(s):  
Base Change ◽  
Molecular Defect ◽  
Fibrinopeptide A ◽  
Fibrin Polymerization ◽  
Prolonged Incubation ◽  
Fibrinogen Binding ◽  
Sds Page ◽  
History Of ◽  
Single Base Change ◽  
Fibrin Monomers

Congenital homozygous dysfibrinogenemia was diagnosed in a man with a history of 2 thrombotic strokes before age 30. His hemostatic profile was characterized by a dramatically prolonged plasma thrombin clotting time, and no clotting was observed with reptilase. Complete clotting of the abnormal fibrinogen occurred after a prolonged incubation of plasma with thrombin. The release of fibrinopeptides A and B by thrombin and of fibrinopeptide A by reptilase were both normal. Thrombin-induced fibrin polymerization was impaired, and no polymerization occurred with reptilase. The polymerization defect was characterized by a defective site “a,” resulting in an absence of interaction between sites A and a, indicated by the lack of fragment D1 (or fibrinogen) binding to normal fibrin monomers depleted in fibrinopeptide A only (Des-AA fm). By SDS-PAGE, the defect was detected on the γ-chain and in its fragment D1. The molecular defect determined by analysis of genomic DNA showed a single base change (A→T) in exon VIII of the γ-chain. The resulting change in the amino acid structure is γ 330 aspartic acid (GAT) → valine (GTT). It is concluded that the residue γ-Asp330 is essential for the normal functioning of the polymerization site a on the fibrinogen γ-chain.

Elektrophoretische Untersuchungen an Mutanten des Phagen Φ X 174

1963 ◽  
Vol 18 (4) ◽  
pp. 290-293 ◽  
Author(s):  
H. G. Aach
Keyword(s):  
Amino Acid ◽  
Mutant Strain ◽  
Spontaneous Mutation ◽  
The Other ◽  
Electrophoretic Mobilities ◽  
The Difference ◽  
Ph Range

The electrophoretic mobilities of the small phages S 13. Φ Χ 174 and of a series of sequential mutants of the latter have been estimated, so that a pH-mobility diagram for each strain could be constructed. The mutants could be separated into two groups by their electrokinetic behaviour, one of the groups reacting more basically over the whole pH-range (3.5 — 9,6) than the other. Parallel with the second spontaneous mutation the mutant strain changed from the first to the second mobility curve. Only one of the following five mutants jumped back to the former curve. It is assumed that the difference in the electrokinetic behaviour is caused by the exchange of nn acidic for an uncharged amino acid and vice versa within the repeating subunit of the protein cover of the phages.

Hereditary Hypodysfibrinogenemia with Defective Release of Fibrinogenopeptide A (Fibrinogen Freiburg)

1979 ◽  
Author(s):  
D. Böttcher ◽  
K. Hasler ◽  
E. Köttgen ◽  
J. Maurath
Keyword(s):  
Bleeding Episode ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Thromboembolic Disease ◽  
Thrombin Time ◽  
Excessive Bleeding ◽  
Major Bleeding Episode ◽  
History Of ◽  
Fibrin Monomers ◽  
Major Defect

A new autoaomally inherited hypodysfibrinogenemia was recognized in four members in three different generations of a family. Only one patient had a major bleeding episode after trauma, the other affected members had no history of excessive bleeding or thromboembolic disease.The thrombin time and Reptilase time of plasma were greatly prolonged and partially corrected by the addition of calcium. Patient plasma prolonged the thrombin time of normal plasma. Fibrinogen levels ranged between 10 to 20 mg/100ml when measured as thrombin-clottable protein, whereas immunologically the fibrinogen levels were only slightly reduced.Functionally the major defect was impaired release of fibrinogenopeptide A upon incubation of the purified abnormal fibrinogen (94% clottable protein) with thrombin and Reptilase. The abnormal fibrinogen showed a delayed polymerisation of its purified fibrin monomers. The described abnormal fibrinogen was indistinguishable from normal fibrinogen by polyacrylamide gel electrophoresis with and without sodium dodecyl sulfate.

TWO ABNORMAL FIBRINOGENS DESIGNATED AS OSAKA II AND MORIOKA WITH A HITHERTO UNIDENTIFIED AMINO ACID SUBSTITUTION; γARG-275 BY CYS

1987 ◽  
Author(s):  
S Terukina ◽  
M Matsuda ◽  
N Yoshida ◽  
K Yamazumi ◽  
Y Takeda ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Substitution ◽  
Sequence Data ◽  
Total Amino Acid ◽  
Reverse Phase Hplc ◽  
Reverse Phase ◽  
Functional Studies ◽  
Sds Page ◽  
Two Populations ◽  
Fibrin Monomers

A hitherto unidentified amino acid substitution of γ Arg-275 by Cys has been found in two abnormal fibrinogens, Osaka II and Morioka. The propositi are both asymptomatic heterozygotes for the abnormality characterized by altered polymerization of fibrin monomers. Reducing SDS-PAGE revealed that fibrinogens derived from thé propositi both consist of two populations; one with a normal and the other with an abnormal longer γ-chain by 0.5 Kd.The γ-γ cross-linking took place nearly normally, however. Analyzing plasmic digests of fibrinogen by SDS-PAGE, we located the abnormality residing in the γ-chain remnant of fragment D. Chromatofocusing of D1 obtained by plasmic digestion in 5 mM Ca++ of purified fibrinogen separated the variant D1 (vD1) from the normal one (nD1) distinctly, as confirmed by SDS-PAGE and functional studies. As anticipated, vD1 failed to interfere with normal fibrin polymerization and thrombin clotting of normal fibrinogen, whereas nD1 inhibited these reactions significantly. After reduction and pyridylethylation, vD1 and nD1 were individually digested with lysylendopeptidase (lysEP). Analyzing the digests by reverse phase HPLC, we noted a single peak present in the digests of vD1 but missing in those of nD1, and vice versa. Analysis of N-terminal five cycles of these peptides suggested that both of them corresponded to the peptide with residues 274302 based on the known sequence data. Primary sequence and total amino acid analyses revealed that γ Arg-275 has been substituted by Cys in both of these abnormal fibrinogens. Analysis of the lysEP-digests of the isolated γ-chain also gave the same result. Since no free SH has been identified at the γ Cys-275 substitute, the variant γ-chain may be endowed with some additive by an S-S linkage. Even if so, elucidation of an apparent elongation by SDS-PAGE of the γ-chain variant must await further investigation. In any case, however, the substitution of γ Arg-275 by Cys may have induced critical alterations in the γ-chain-dependent polymerization site in the D domain in these two abnormal fibrinogens.

PROTHROMBIN TOKUSHIMA: A REPLACEMENT OF ARGININE-418 BY TRYPTOPHAN THAT IMPAIRS THE FIBRINOGEN CLOTTING ACTIVITY OF DERIVED THROMBIN TOKUSHIMA

1987 ◽  
Author(s):  
T Miyata ◽  
T Morita ◽  
T Inomoto ◽  
S Kawauchi ◽  
A Shirakami ◽  
...  
Keyword(s):  
Amino Acid ◽  
Factor Xa ◽  
Recurrent Bleeding ◽  
Bleeding Tendency ◽  
Japanese Family ◽  
Single Nucleotide Change ◽  
Lysyl Endopeptidase ◽  
Normal Enzyme ◽  
The Difference ◽  
Aggregation Activity

Recently, we identified a Japanese family having an abnormal prothrombin, designated “prothrombin Tokushima”. The propositus of this family is a 10-year-old female suffering from bleeding tendency. She is a double heterozygote transmitted from both the maternal dysprothrombinemia and the paternal hypoprothrombinemia. The purified prothrombin Tokushima exhibited the following properties: (i) factor Xa-catalyzed proteolysis of the abnormal prothrombin in the presence of factor Va, phospholipids and calcium ions yielded a two-chain thrombin, fragment 1 and fragment 2, (ii) thrombin derived from the abnormal molecule exhibited 21.5 % clotting activity relative to normal thrombin and reduced platelet aggregation activity, (iii) the apparent Km of thrombin Tokushima towards Boc-Val-Pro-Arg-MCA was 8-fold larger than that of normal thrombin, and the kcat value was approximately one-half of that of the normal enzyme, and (iv) its active site was fully titrable using P-nitrophenyl-P’-guanidinobenzoate.Subsequently, the structural studies on the abnormal thrombin were performed to identify the difference responsible for its reduced fibrinogen clotting activity upon conversion to thrombin. Amino acid sequence analysis of a peptide isolated from a lysyl endopeptidase digest of the abnormal thrombin indicated that Arg-418 (equivalent to Asn-101 in the chymotrypsin numbering system) had been replaced by Trp. This amino acid substitution can result from a single nucleotide change in the codon for Arg→4l8 (CGG→TGG). The Arg→Trp replacement found in the thrombin portion of prothrombin Tokushima appears to reduce its interaction with various substrates including fibrinogen and platelet receptors, and accounts for the recurrent bleeding episode observed in the propositus.

Isolation and Structural Charaderization of a Potent Inhibitor of Coagulation Factor Xa from the Leech Haementeria ghilianii

1989 ◽  
Vol 61 (03) ◽  
pp. 437-441 ◽  
Author(s):  
Cindra Condra ◽  
Elka Nutt ◽  
Christopher J Petroski ◽  
Ellen Simpson ◽  
P A Friedman ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Salivary Gland ◽  
Amino Acid ◽  
Amino Acid Sequence ◽  
Western Blot ◽  
Potent Inhibitor ◽  
Factor Xa ◽  
Coagulation Factor ◽  
Sds Page ◽  
Bovine Factor

SummaryThe present work reports the discovery and charactenzation of an anticoagulant protein in the salivary gland of the giant bloodsucking leech, H. ghilianii, which is a specific and potent inhibitor of coagulation factor Xa. The inhibitor, purified to homogeneity, displayed subnanomolar inhibition of bovine factor Xa and had a molecular weight of approximately 15,000 as deduced by denaturing SDS-PAGE. The amino acid sequence of the first 43 residues of the H. ghilianii derived inhibitor displayed a striking homology to antistasin, the recently described subnanomolar inhibitor of factor Xa isolated from the Mexican leech, H. officinalis. Antisera prepared to antistasin cross-reacted with the H. ghilianii protein in Western Blot analysis. These data indicate that the giant Amazonian leech, H. ghilianii, and the smaller Mexican leech, H. officinalrs, have similar proteins which disrupt the normal hemostatic clotting mechanisms in their mammalian host’s blood.

Sign in / Sign up

Export Citation Format

Share Document