A Simple and Practical Method for Isolation of an Early Plasmin Degradation Product of Human Fibrinogen

The stability of disodium sulfobenzylpenicillin (SB-PC) in several large-volume parenteral solutions was studied. A study was also made of the influences of other additive drugs upon the stability of SB-PC in SB-PC IV fluid admixtures. Quantitative determinations were made by iodometry of the residual rate over time of SB-PC. The residual rate of SB-PC in 5-fluorouracil (5-FU injection)-10 percent glucose admixture was 67 percent after 24 hours at 25 °C, (77 °F) and in aminophylline (aminophylline injection J.P.)-10 percent glucose admixture it was 54 percent after 24 hours. It was found that the decomposition of SB-PC was due to amines which were added to these injections. In an admixture of amino acid and SB-PC, the residual rate of SB-PC decreased linearly with the increase in amino acid concentration. In the 12 kinds of amino acids used in this study, the greatest decrease was observed in the admixture with lysine, the residual rate being 76.7 percent. For the study of degradation products of SB-PC, thin layer chromatography was employed and a spot of the degradation product was detected at an Rf value of 0.2.

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Limited Proteolysis of the Purified Aα Chain of Human Fibrinosen by Plasmin

10.1055/s-0039-1689532 ◽

1975 ◽

Author(s):

L. Williams ◽

G. Murano

Keyword(s):

Molecular Weight ◽

Amino Acid ◽

Peptide Mapping ◽

Proteolytic Enzymes ◽

Degradation Products ◽

Limited Proteolysis ◽

Gel Filtration ◽

Terminal Region ◽

Low Molecular Weight ◽

Human Fibrinogen

Based on evidence that a portion of circulating fibrinogen consists of a family of catabolic intermediates formed by proteolytic degradation of the COOH terminal region of Aα chains, we attempted to obtain early degradation products using the purified alkylated Aα chain derivative of human fibrinogen as the substrate and plasmin as the enzyme. Having established optimal conditions, a preparative quantity of material was digested in 0.1 M tris buffer pH = 9.5; time = 4 min; E/S ratio = 1/75 (mole/mole); temp = 37° C. Low molecular weight fragments were separated from the larger species, and further purified by gel filtration on Sephadex G-100. Selected early fragments were analyzed by polycrylamide gel electrophoresis, amino acid composition, peptide mapping and partial N-terminal amino acid sequence. Two of the earliest low molecular weight fragments released by plasmin were derived from the N-terminal region of the Aα chain. Their molecular size was estimated at about 10,000 daltons. One fragment contains fibrinopeptide A; both fragments extend beyond Met-51. Our data indicate that: a) the specificity of plasmin on the purified Aα chain differs from that on intact fibrinogen; or b) proteolytic enzymes other than or in addition to plasmin are responsible for the formation of early catabolic fibrinogen intermediates having a degraded Aα chain.(Supported by USPHS N. I. H. Grant HL 14142.)

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Permeability Enhancing and Chemotactic Activities of Lower Molecular Weight Degradation Products of Human Fibrinogen

Thrombosis and Haemostasis ◽

10.1055/s-0038-1650136 ◽

1981 ◽

Vol 45 (01) ◽

pp. 090-094 ◽

Cited By ~ 52

Author(s):

Katsuo Sueishi ◽

Shigeru Nanno ◽

Kenzo Tanaka

Keyword(s):

Molecular Weight ◽

Degradation Products ◽

Electron Microscopic Observation ◽

Chemotactic Activity ◽

Lower Molecular Weight ◽

Electron Microscopic ◽

Human Fibrinogen ◽

Rabbit Skin ◽

Molecular Weights ◽

Active Fragments

SummaryFibrinogen degradation products were investigated for leukocyte chemotactic activity and for enhancement of vascular permeability. Both activities increased progressively with plasmin digestion of fibrinogen. Active fragments were partially purified from 24 hr-plasmin digests. Molecular weights of the permeability increasing and chemotactic activity fractions were 25,000-15,000 and 25,000 respectively. Both fractions had much higher activities than the fragment X, Y, D or E. Electron microscopic observation of the small blood vessels in rabbit skin correlated increased permeability with the formation of characteristic gaps between adjoining endothelial cells and their contraction.These findings suggest that lower molecular weight degradation products of fibrinogen may be influential in contributing to granulocytic infiltration and enhanced permeability in lesions characterized by deposits of fibrin and/or fibrinogen.

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Complete Covalent Structure of Human Platelet Factor 4

Thrombosis and Haemostasis ◽

10.1055/s-0038-1657071 ◽

1979 ◽

Vol 42 (05) ◽

pp. 1652-1660 ◽

Cited By ~ 4

Author(s):

Francis J Morgan ◽

Geoffrey S Begg ◽

Colin N Chesterman

Keyword(s):

Amino Acids ◽

Molecular Weight ◽

Amino Acid ◽

Amino Acid Sequence ◽

Human Platelet ◽

Platelet Factor 4 ◽

Platelet Factor ◽

Disulphide Bonds ◽

Covalent Structure

SummaryThe amino acid sequence of the subunit of human platelet factor 4 has been determined. Human platelet factor 4 consists of identical subunits containing 70 amino acids, each with a molecular weight of 7,756. The molecule contains no methionine, phenylalanine or tryptophan. The proposed amino acid sequence of PF4 is: Glu-Ala-Glu-Glu-Asp-Gly-Asp-Leu-Gln-Cys-Leu-Cys-Val-Lys-Thr-Thr-Ser- Gln-Val-Arg-Pro-Arg-His-Ile-Thr-Ser-Leu-Glu-Val-Ile-Lys-Ala-Gly-Pro-His-Cys-Pro-Thr-Ala-Gin- Leu-Ile-Ala-Thr-Leu-Lys-Asn-Gly-Arg-Lys-Ile-Cys-Leu-Asp-Leu-Gln-Ala-Pro-Leu-Tyr-Lys-Lys- Ile-Ile-Lys-Lys-Leu-Leu-Glu-Ser. From consideration of the homology with p-thromboglobulin, disulphide bonds between residues 10 and 36 and between residues 12 and 52 can be inferred.

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Characterization of an isozyme of S-adenosylmethionine synthetase from rat liver

Canadian Journal of Biochemistry and Cell Biology ◽

10.1139/o84-038 ◽

1984 ◽

Vol 62 (5) ◽

pp. 276-279 ◽

Cited By ~ 1

Author(s):

C. H. Lin ◽

W. Chung ◽

K. P. Strickland ◽

A. J. Hudson

Keyword(s):

Amino Acids ◽

Molecular Weight ◽

Amino Acid ◽

Enzymatic Synthesis ◽

Gel Filtration ◽

Deae Cellulose ◽

Aromatic Amino Acids ◽

Adenosylmethionine Synthetase ◽

Cellulose Column

An isozyme of S-adenosylmethionine synthetase has been purified to homogeneity by ammonium sulfate fractionation, DEAE-cellulose column chromatography, and gel filtration on a Sephadex G-200 column. The purified enzyme is very unstable and has a molecular weight of 120 000 consisting of two identical subunits. Amino acid analysis on the purified enzyme showed glycine, glutamate, and aspartate to be the most abundant and the aromatic amino acids to be the least abundant. It possesses tripolyphosphatase activity which can be stimulated five to six times by S-adenosylmethionine (20–40 μM). The findings support the conclusion that an enzyme-bound tripolyphosphate is an obligatory intermediate in the enzymatic synthesis of S-adenosylmethionine from ATP and methionine.

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The C-terminal sequence of amino acid residues of κ-casein isolated from buffalo's milk

Journal of Dairy Research ◽

10.1017/s0022029900012176 ◽

1967 ◽

Vol 34 (1) ◽

pp. 85-88 ◽

Cited By ~ 4

Author(s):

M. H. Abd El-Salam ◽

W. Manson

Keyword(s):

Amino Acids ◽

Molecular Weight ◽

Amino Acid ◽

Peptide Chain ◽

Amino Acid Residues ◽

Terminal Sequence ◽

Carboxypeptidase A ◽

Phosphorous Content ◽

Single Peptide

SummaryWhen κ-casein from buffalo's milk was treated with carboxypeptidase A (EC 3. 4. 2. 1),4 amino acids, valine, threonine, serine and alanine were released from the protein in a manner consistent with the view that they originate in the C-terminal sequence of a single peptide chain. The amounts produced suggest a minimum molecular weight for buffalo κ-casein of approximately 17000, in agreement with the value calculated from the phosphorous content on the basis of the presence of 2 phosphorus atoms/molecule. A comparison is made with the C-terminal sequence reported for bovine κ-casein.

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Isolation and characterization of some novel genes of the apolipoprotein A-I family in Japanese eel, Anguilla japonica

Open Life Sciences ◽

10.2478/s11535-011-0042-8 ◽

2011 ◽

Vol 6 (4) ◽

pp. 545-557 ◽

Cited By ~ 2

Author(s):

Malay Choudhury ◽

Takahiro Oku ◽

Shoji Yamada ◽

Masaharu Komatsu ◽

Keita Kudoh ◽

...

Keyword(s):

Amino Acids ◽

Molecular Weight ◽

Amino Acid ◽

Consensus Sequence ◽

Structural Features ◽

Lipid Binding ◽

Japanese Eel ◽

Amino Acid Sequences ◽

Novel Genes ◽

Isolation And Characterization

AbstractApolipoproteins such as apolipoprotein (apo) A-I, apoA-IV, and apoE are lipid binding proteins synthesized mainly in the liver and the intestine and play an important role in the transfer of exogenous or endogenous lipids through the circulatory system. To investigate the mechanism of lipid transport in fish, we have isolated some novel genes of the apoA-I family, apoIA-I (apoA-I isoform) 1–11, from Japanese eel by PCR amplification. Some of the isolated genes of apoIA-I corresponded to 28kDa-1 cDNAs which had already been deposited into the database and encoded an apolipoprotein with molecular weight of 28 kDa in the LDL, whereas others seemed to be novel genes. The structural organization of all apoIA-Is consisted of four exons separated by three introns. ApoIA-I10 had a total length of 3232 bp, whereas other genes except for apoIA-I9 ranged from 1280 to 1441 bp. The sequences of apoIA-Is at the exon-intron junctions were mostly consistent with the consensus sequence (GT/AG) at exon-intron boundaries, whereas the sequences of 3′ splice acceptor in intron 1 of apoIA-I1-7 were (AC) but not (AG). The deduced amino acid sequences of all apoIA-Is contained a putative signal peptide and a propeptide of 17 and 5 amino acid residues, respectively. The mature proteins of apoIA-I1-3, 7, and 8 consisted of 237 amino acids, whereas those of apoIA-I4-6 consisted of 239 amino acids. The mature apoIA-I10 sequence showed 65% identity to amino acid sequence of apoIA-I11 which was associated with an apolipoprotein with molecular weight of 23 kDa in the VLDL. All these mature apoIA-I sequences satisfied the common structural features depicted for the exchangeable apolipoproteins such as apoA-I, apoA-IV, and apoE but apoIA-I11 lacked internal repeats 7, 8, and 9 when compared with other members of apoA-I family. Phylogenetic analysis showed that these novel apoIA-Is isolated from Japanese eel were much closer to apoA-I than apoA-IV and apoE, suggesting new members of the apoA-I family.

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Molecular analysis of nuc-1+, a gene controlling phosphorus acquisition in Neurospora crassa

Molecular and Cellular Biology ◽

10.1128/mcb.10.11.5839-5848.1990 ◽

1990 ◽

Vol 10 (11) ◽

pp. 5839-5848

Author(s):

S Kang ◽

R L Metzenberg

Keyword(s):

Amino Acids ◽

Molecular Weight ◽

Amino Acid ◽

Amino Acid Sequence ◽

Neurospora Crassa ◽

Phosphorus Acquisition ◽

Regulatory Factor ◽

Physical Interactions ◽

Negative Regulatory Factor ◽

High Phosphate

In response to phosphorus starvation, Neurospora crassa makes several enzymes that are undetectable or barely detectable in phosphate-sufficient cultures. The nuc-1+ gene, whose product regulates the synthesis of these enzymes, was cloned and sequenced. The nuc-1+ gene encodes a protein of 824 amino acids with a predicted molecular weight of 87,429. The amino acid sequence shows homology with two yeast proteins whose functions are analogous to that of the NUC-1 protein. Two nuc-1+ transcripts of 3.2 and 3.0 kilobases were detected; they were present in similar amounts during growth at low or high phosphate concentrations. The nuc-2+ gene encodes a product normally required for NUC-1 function, and yet a nuc-2 mutation can be complemented by overexpression of the nuc-1+ gene. This implies physical interactions between NUC-1 protein and the negative regulatory factor(s) PREG and/or PGOV. Analysis of nuc-2 and nuc-1; nuc-2 strains transformed by the nuc-1+ gene suggests that phosphate directly affects the level or activity of the negative regulatory factor(s) controlling phosphorus acquisition.

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DESIGNING BALANCED FEEDS FOR INDUSTRIAL AQUACULTURE USING PROTEIN HYDROLYSATES OF FISH BY-PASS RAW MATERIALS

Fisheries ◽

10.37663/0131-6184-2021-4-81-88 ◽

2021 ◽

Vol 2021 (4) ◽

pp. 81-88

Author(s):

Olga Mezenova ◽

Dmitriy Pyanov ◽

Svetlana Agafonova ◽

Natalia Mezenova ◽

V. Volkov

Keyword(s):

Amino Acids ◽

Molecular Weight ◽

Amino Acid ◽

Raw Materials ◽

Protein Hydrolysates ◽

Total Protein Content ◽

Alternative Sources ◽

Amino Acid Balance ◽

Limiting Amino Acids ◽

By Products

The perspective of the production of domestic compound feed for the development of industrial aquaculture in Russia is shown. Alternative sources of protein in mixed fodder for salmon and sturgeon have been investigated. The advantages of using protein hydrolysates instead of a part of fishmeal in compound feed are described. The advantages of protein hydrolysates from fish by-products are considered, the chemical composition and molecular fractional composition of sublimated protein hydrolysates obtained by enzymatic and thermal pathways from sardinella scales and ridges are studied. The presence in hydrolysates of 53.3 - 97.7% of low molecular weight peptides with a molecular weight of less than 10 kDa with a total protein content of 80.8-94.1% was established. Indicators of amino acid balance (scor) of hydrolyzates of scales and ridges of sardinella were calculated in relation to the established requirements for amino acids in salmonids. Indicators of amino acid balance (scor) of hydrolyzates of scales and ridges of sardinella were calculated in relation to the established requirements for amino acids in salmonids.It was found that the introduction of an enzymatically obtained hydrolyzate is more favorable for an increase in the content of limiting amino acids in mixed feed, and the use of sardinella scales for hydrolysis is more preferable than its ridges.

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Molecular analysis of nuc-1+, a gene controlling phosphorus acquisition in Neurospora crassa.

Molecular and Cellular Biology ◽

10.1128/mcb.10.11.5839 ◽

1990 ◽

Vol 10 (11) ◽

pp. 5839-5848 ◽

Cited By ~ 34

Author(s):

S Kang ◽

R L Metzenberg

Keyword(s):

Amino Acids ◽

Molecular Weight ◽

Amino Acid ◽

Amino Acid Sequence ◽

Neurospora Crassa ◽

Phosphorus Acquisition ◽

Regulatory Factor ◽

Physical Interactions ◽

Negative Regulatory Factor ◽

High Phosphate

In response to phosphorus starvation, Neurospora crassa makes several enzymes that are undetectable or barely detectable in phosphate-sufficient cultures. The nuc-1+ gene, whose product regulates the synthesis of these enzymes, was cloned and sequenced. The nuc-1+ gene encodes a protein of 824 amino acids with a predicted molecular weight of 87,429. The amino acid sequence shows homology with two yeast proteins whose functions are analogous to that of the NUC-1 protein. Two nuc-1+ transcripts of 3.2 and 3.0 kilobases were detected; they were present in similar amounts during growth at low or high phosphate concentrations. The nuc-2+ gene encodes a product normally required for NUC-1 function, and yet a nuc-2 mutation can be complemented by overexpression of the nuc-1+ gene. This implies physical interactions between NUC-1 protein and the negative regulatory factor(s) PREG and/or PGOV. Analysis of nuc-2 and nuc-1; nuc-2 strains transformed by the nuc-1+ gene suggests that phosphate directly affects the level or activity of the negative regulatory factor(s) controlling phosphorus acquisition.

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