scholarly journals Identification of two major proteins of bovine pancreatic stones as immunoreactive forms of trypsinogens

1982 ◽  
Vol 205 (3) ◽  
pp. 543-549 ◽  
Author(s):  
A De Caro ◽  
L Multigner ◽  
H Vérine
Keyword(s):  
Molecular Weight ◽  
Isoelectric Focusing ◽  
Sodium Citrate ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Charge Heterogeneity ◽  
Two Dimensional Electrophoresis ◽  
Pancreatic Stones ◽  
Dimensional Electrophoresis

Two major proteins have been identified in sodium citrate extracts of bovine pancreatic stones from 15 glands with lithiasis. They were found to have a molecular weight of about 24 000 and were further characterized by a variety of methods, including polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate, isoelectric focusing, two-dimensional electrophoresis, immunodiffusion, immunoelectrophoresis and determination of N-terminal residues. These two immunologically and electrophoretically different proteins were definitely shown to be immunoreactive forms of anionic and cationic trypsinogens, which are normal components of pancreatic juice. However, in contrast with both secretory trypsinogens, the stone proteins displayed an important charge heterogeneity under isoelectric-focusing conditions. A possible role for both secretory trypsinogens in pancreatic lithogenesis is suggested by the reproducibility of the data. Finally, two minor proteins with a lower molecular weight (about 11 000-13 000) have also been found to be present in all extracts, but have not yet been identified.

scholarly journals Separation and partial characterization of guinea-pig caseins

1978 ◽  
Vol 173 (2) ◽  
pp. 633-641 ◽  
Author(s):  
R K Craig ◽  
D McIlreavy ◽  
R L Hall
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Guinea Pig ◽  
Amino Acid Composition ◽  
Acid Composition ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Gel Filtration ◽  
Exchange Chromatography

1. Guinea-pig caseins A, B and C were purified free of each other by a combination of ion-exchange chromatography and gel filtration. 2. Determination of the amino acid composition showed all three caseins to contain a high proportion of proline and glutamic acid, but no cysteine. This apart, the amino acid composition of the three caseins was markedly different, though calculated divergence values suggest that some homology may exist between caseins A and B. Molecular-weight estimates based on amino acid composition were in good agreement with those based on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. 3. N-Terminal analysis showed lysine, methionine and lysine to be the N-terminal residues of caseins A, B and C respectively. 4. Two-dimensional separation of tryptic digests revealed a distinctive pattern for each casein. 5. All caseins were shown to be phosphoproteins. The casein C preparation also contained significant amounts of sialic acid, neutral and amino sugars. 6. The results suggest that each casein represents a separate gene product, and that the low-molecular-weight proteins are not the result of a post-translational cleavage of the largest. All were distinctly different from the whey protein alpha-lactalbumin.

scholarly journals Haemoglobin from the tadpole shrimp, Lepidurus apus lubbocki Characterization of the molecule and determination of the number of polypeptide chains

1979 ◽  
Vol 183 (2) ◽  
pp. 325-330 ◽  
Author(s):  
E Ilan ◽  
E Daniel
Keyword(s):  
Molecular Weight ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Sedimentation Coefficient ◽  
Sedimentation Equilibrium ◽  
Guanidinium Chloride ◽  
Tadpole Shrimp ◽  
Polypeptide Chains

Haemoglobin from the tadpole shrimp, Lepidurus apus lubbocki, was found to have a sedimentation coefficient (s020,w) of 19.3 +/- 0.2 S and a molecular weight, as determined by sedimentation equilibrium, of 798000 +/- 20000. The amino acid composition showed the lack of cysteine and cystine residues. A haem content of 3.55 +/- 0.03% was determined, corresponding to a minimal mol.wt. of 17400 +/- 200. The pH-independence in the range pH 5-11 of the sedimentation coefficient indicates a relatively high stability of the native molecule. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis gave one band with mobility corresponding to a mol.wt. of 34000 +/- 1500. The molecular weight of the polypeptide chain was determined to be 32800 +/- 800 by sedimentation equilibrium in 6 M-guanidinium chloride and 0.1 M-2-mercaptoethanol. The findings indicate that Lepidurus haemoglobin is composed of 24 identical polypeptide chains, carrying two haem groups each.

Comparative study on polypeptide patterns of larvae of Trichinella isolates by two-dimensional electrophoresis

1987 ◽  
Vol 61 (3) ◽  
pp. 225-228 ◽  
Author(s):  
T. Garate ◽  
L. Rivas
Keyword(s):  
Comparative Study ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Polypeptide Patterns ◽  
Muscle Larvae ◽  
The Comparative Study ◽  
Dimensional Electrophoresis

ABSTRACTThe two-dimensional patterns (isoelectrofocusing-IEF/polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate-SDS) of S3 fractions of muscle larvae of four Trichinella isolates were compared. The comparative study concerned six groups of polypeptides. It was observed that the Garkavi isolate of Trichinella pseudospiralis was clearly different from the other isolates, and it showed the simplest IEF/SDS polypeptide pattern. The C-76 isolate of T. nelsoni had only four of the six groups, distinguishing it from the GM-1 isolate of T. spiralis and the Boev isolate of T. nativa that showed all the indicated groups.

PURIFICATION AND PROPERTIES OF CHORIONIC GONADOTROPHIN FROM TROPHOBLASTIC TISSUE, URINE AND PLASMA OF A PATIENT WITH A HYDATIDIFORM MOLE

1973 ◽  
Vol 56 (3) ◽  
pp. 441-NP ◽  
Author(s):  
A. PALA ◽  
M. MEIRINHO ◽  
G. BENAGIANO
Keyword(s):  
Molecular Weight ◽  
Isoelectric Focusing ◽  
Sodium Dodecyl Sulphate ◽  
Polyacrylamide Gel Electrophoresis ◽  
Hydatidiform Mole ◽  
Sodium Dodecyl ◽  
Deae Cellulose ◽  
Chorionic Gonadotrophin ◽  
Tissue Preparation ◽  
Trophoblastic Tissue

SUMMARY Purified human chorionic gonadotrophin (HCG) fractions were obtained from urine, peripheral plasma and chorionic tissue of a woman with hydatidiform mole by a combination of DEAE-cellulose and DEAE-Sephadex ion exchange chromatography and gel filtration on Sephadex G-100. Homogeneity of the purified fractions obtained was demonstrated by analytical polyacrylamide gel electrophoresis and immunoelectrophoresis. The preparation from plasma had a higher ratio of biological to immunological activity than the urine or tissue preparation. The biological potency of the plasma HCG was 23000 i.u./mg, whereas the urinary and the trophoblastic tissue preparations showed a potency of 20000 and 6500 i.u./mg, respectively. Further characterization of the three fractions was achieved by analytical isoelectric focusing and determination of the molecular weight. The peak of radioimmunological activity after isoelectric focusing was found to be, in all three cases, at pH 4·90 and the molecular weight obtained after incubation with sodium dodecyl sulphate and urea was around 63000 When the molecular weight was determined after incubation in a medium containing 4 m-urea, sodium dodecyl sulphate and 2-mercaptoethanol two separate fractions were obtained from each of the three preparations. The heavier fraction showed a molecular weight of approximately 35000 and the lighter fraction had a molecular weight of approximately 27000.

scholarly journals PURIFICATION AND MOLECULAR WEIGHT DETERMINATION OF KERATINASE ISOLATED FROM STREPTOMYCES MALAYSIENSIS

2017 ◽  
Vol 9 (10) ◽  
pp. 154
Author(s):  
M. Pavani ◽  
G. Girijasankar ◽  
K. Mallika ◽  
G. Vidhya Sagar
Keyword(s):  
Molecular Weight ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Sole Source ◽  
Molecular Weight Determination ◽  
Termite Mound ◽  
Phylogenetic Tree Analysis ◽  
Sds Page ◽  
Streptomyces Malaysiensis

Objective: The aim of the present study was to purify and determine the molecular weight of keratinase isolated from Streptomyces malaysiensis.Methods: For that purpose purification was done using ammonium sulphate and Sephadex-LH 100 column chromatography. Further, the fractions were pooled and subjected to molecular weight determination using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE).Results: The obtained results showed keratinase with 47.57% recovery, 3.5-fold purification and an estimated molecular mass of 27,000 Da. Keratinase showed an optimal activity at 60 οC and pH 8. Keratinase activity of the purified product was assayed with feather powder as a substrate. The isolated strain was identified as Streptomyces malaysiensis based on phylogenetic tree analysis. The strain isolated from termite mound soil showed the highest keratinase activity, which could be considered a microorganism of environmental origin.Conclusion: The production of keratinase on simple media with feathers as sole source allowing its production from the cheap substrate and a commercial production with low production cost. Stability in the presence of detergents, surfactants and solvents make this keratinase extremely useful for a biotechnological process involving keratin.

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