Studies on Fibrinopeptides from Mammals.

Blombäck, Birger; Blombäck, Margareta; Gröndahl, Nils Jakob; Enari, T.-M.

doi:10.3891/acta.chem.scand.19-1789

Cited by 47 publications

(17 citation statements)

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“…Nucleotides 25-54 code for 10 amino acids which are identical to residues 44-53 of the 83 chain of bovine fibrinogen (31 530 540 550 560 570 580 590 600 240 250 260 Pro reported amino-terminal sequence of the p chain of bovine firesidues in corresponding positions being identical. The posibrinogen (30,31,33), the complete amino acid sequence of the tions of all the cystines and tryptophans are identical. The amide ,B chain of bovine fibrinogen has been constructed (Fig.…”

mentioning

confidence: 90%

Characterization of a cDNA clone coding for the beta chain of bovine fibrinogen.

Chung¹,

Rixon²,

MacGillivray³

et al. 1981

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

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Recombinant plasmids containing bovine cDNA have been screened with a radiolabeled cDNA enriched for bovine fibrinogen. A number of plasmids containing cDNAs for fibrinogen were identified by this assay. One 66,124, 52,306, and 46,474, respectively (4). During the coagulation process, thrombin removes fibrinopeptides A and B from the amino termini of the a and 13 chains, respectively. This leads to the formation of fibrin monomers which are subsequently polymerized and crosslinked to form a tough, insoluble, fibrin clot (5,6).The amino acid sequences of the a, 13, and y chains of human fibrinogen have been determined (7-11). A high degree of homology among the three chains suggests that they may have been derived from a common ancestral gene (11). A distended polydomainal structure linked by interdomainal supercoiled ahelices has been proposed (12-15).Fibrinogen is synthesized in hepatic parenchymal cells (16) where the a, 13, and 'y chains are synthesized from individual mRNAs (17). Little is known, however, about the subsequent processing, assembly, and secretion of this protein into the plasma. The biosynthesis of fibrinogen is stimulated during inflammation, injury, or bacterial infection, and this can lead to a substantial increase in its plasma level (18). The mechanism for this increase has not been established. A number of questions regarding the structure, function, and biosynthesis of fibrinogen can be examined with cDNA probes to specific mRNAs. Accordingly, we have developed methods for the isolation and characterization of recombinant plasmids containing cDNA coding for the three chains of this protein.In the present report, we describe the properties of a plasmid containing a cDNA coding for the ,B chain. MATERIALS AND METHODSEnzymes. The Klenow fragment of Escherichia coli polymerase I and T4 polynucleotide kinase were purchased from P-L Biochemicals. All restriction endonucleases were purchased from Bethesda Research Laboratories (Rockville, MD), except Sau 3A which was purchased from New England BioLabs. Bovine fibrinogen was purchased from Calbiochem.Construction of Recombinant Plasmids. Reverse transcription of enriched bovine liver mRNA, synthesis of doublestranded cDNA, homopolymeric G/C tailing, construction of recombinant plasmids by using pBR322 as vector, and cloning into E. coli K-12, strain RRI, have been reported (19). The cloning was performed under P3-EK1 conditions in compliance with the revised (1978) guidelines for recombinant DNA research from the National Institutes of Health.Screening of Recombinant Plasmids. Bovine liver polysomes were prepared as described (17,20). mRNA for fibrinogen was enriched by immunoadsorption with Staphylococcus aureus cells and affinity-purified antibodies to bovine fibrinogen (21). Radiolabeled cDNA was prepared from the enriched mRNA and was used as a probe in a solution hybridization screening procedure as described (19).Restriction Endonuclease Mapping. The locations of restriction enzyme sites on the inserted DNA were determined by ...

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confidence: 90%

Characterization of a cDNA clone coding for the beta chain of bovine fibrinogen.

Chung¹,

Rixon²,

MacGillivray³

et al. 1981

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

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“…In the rabbit sequence of the N-terminal region corresponding to FPA (A· 1-16) or fibrinopeptide B (FPB; Bß 1-13), only a few amino acid residues (e.g., Phe 8 and Arg 16 in FPA, Arg 13 in FPB) were homologous between rabbit and human or bovine sequences. The variability of FPA and FPB sequences among mammalian species had been reported by Blombäck et al [25,26]. On the contrary, the N-termini of fibrin ·-, ß-, and Á-chains were highly homologous between rabbit and human or bovine fibrins, suggesting that the structure of N-terminal fragments of rabbit fibrin may be analogous to human and bovine fibrin.…”

Section: Discussionmentioning

confidence: 87%

Bound Thrombin from Crushed Clots Is Composed of α-Thrombin and the N-Terminal Regions of α- and γ-Chains of Fibrinogen

Kinjoh

Nakamura²,

Zeng³

et al. 2002

Pathophysiol Haemos Thromb

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We aimed at clarifying the structural characteristics of the bound thrombin that is liberated by mechanical breakdown of fibrin clots. Fibrin clots were prepared with bovine thrombin and rabbit fibrinogen, and were crushed mechanically with a glass rod. The supernatant of the crushed clots was subjected to immunoaffinity chromatography to isolate the bound thrombin. Western blotting analysis revealed that the bound thrombin could be reacted with both antithrombin and antifibrinogen under unreduced conditions. SDS-PAGE under reduced conditions revealed that there were three bands, two of which were found to be the N-terminal fragments of the α- and γ-chains of fibrinogen. The bound thrombin could be dissociated into three distinct fibrin fragments and bovine α-thrombin when denatured by 8 M urea. Thus, the bound thrombin liberated from crushed clots is a stable complex between bovine α-thrombin and fibrin fragments of the N-terminal regions of rabbit α- and γ-chains.

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“…They 23 Tuppy, 1958. The conference was organized by the Gesellschaft Deutscher Naturforscher und Ä rtze, The results of yeast cytochrome-c are presented in Tuppy and Dus, 1958. 24 Blomba¨ck et al, 1965Blomba¨ck et al, , p. 1789 Anfinsen et al, 1959Anfinsen et al, , p. 1118. 26 Anfinsen, 1959, p. 143. focused on structural similarities, assuming that natural selection had eliminated structural variations in functionally important regions of proteins.…”

Section: Comparing Sequences To Understand Protein Functionmentioning

confidence: 97%

Collecting, Comparing, and Computing Sequences: The Making of Margaret O. Dayhoff’s Atlas of Protein Sequence and Structure, 1954–1965

Strasser

2009

J Hist Biol

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Collecting, comparing, and computing molecular sequences are among the most prevalent practices in contemporary biological research. They represent a specific way of producing knowledge. This paper explores the historical development of these practices, focusing on the work of Margaret O. Dayhoff, Richard V. Eck, and Robert S. Ledley, who produced the first computer-based collection of protein sequences, published in book format in 1965 as the Atlas of Protein Sequence and Structure. While these practices are generally associated with the rise of molecular evolution in the 1960s, this paper shows that they grew out of research agendas from the previous decade, including the biochemical investigation of the relations between the structures and function of proteins and the theoretical attempt to decipher the genetic code. It also shows how computers became essential for the handling and analysis of sequence data. Finally, this paper reflects on the relationships between experimenting and collecting as two distinct "ways of knowing" that were essential for the transformation of the life sciences in the twentieth century.

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Studies on Fibrinopeptides from Mammals.

Cited by 47 publications

References 0 publications

Characterization of a cDNA clone coding for the beta chain of bovine fibrinogen.

Characterization of a cDNA clone coding for the beta chain of bovine fibrinogen.

Bound Thrombin from Crushed Clots Is Composed of α-Thrombin and the N-Terminal Regions of α- and γ-Chains of Fibrinogen

Collecting, Comparing, and Computing Sequences: The Making of Margaret O. Dayhoff’s Atlas of Protein Sequence and Structure, 1954–1965

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