scholarly journals The amino acid sequence of dromedary pancreatic ribonuclease.

1975 ◽  
Vol 147 (3) ◽  
pp. 505-511 ◽  
Author(s):  
G W Welling ◽  
G Groen ◽  
J J Beintema
Keyword(s):  
Amino Acids ◽  
Polypeptide Chain ◽  
Proteolytic Enzymes ◽  
Gel Filtration ◽  
Paper Electrophoresis ◽  
Pancreatic Tissue ◽  
British Library ◽  
Peptide Bonds ◽  
Pancreatic Ribonuclease ◽  
External Loop

Dromedary (Camelus dromedarius) RNAase (ribonuclease) was isolated from pancreatic tissue by affinity chromatography. Peptides obtained by digestion with different proteolytic enzymes and CNBr were isolated by gel filtration, preparative high-voltage paper electrophoresis and paper chromatography. Peptides were sequenced by the dansyl-Edman method. All peptide bonds were overlapped by one or more peptides. The polypeptide chain consists of 123 amino acids. A deletion (position 39) was observed in an external loop of the polypeptide chain (residues 35-40), as was found earlier to horse RNAase (Scheffer & Beintema, 1974). A heterogeneity was found at position 103 (glutamine and lysine). Dromedary RNAase differs at 23-32% of the positions from all other pancreatic RNAases sequenced to date. In evolutionary terms this indicates that dromedary RNAase has evolved independently during the larger part of the evolution of the mammals. Detailed evidence for the sequence has been deposited as Supplementary Publication SUP 50046 (14 pages) at the British Library (Lending Division), Boston Spa, Wetherby, W. Yorks. LS23 7BQ, U.K., from whom copies may be obtained on the terms given in Biochem. J. (1975) 145, 5.

scholarly journals The amino acid sequence of pike-whale (lesser-rorqual) pancreatic ribonuclease

1976 ◽  
Vol 157 (2) ◽  
pp. 317-323 ◽  
Author(s):  
M Emmens ◽  
G W Welling ◽  
J J Beintema
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
High Voltage ◽  
Proteolytic Enzymes ◽  
Gel Filtration ◽  
Paper Electrophoresis ◽  
British Library ◽  
Balaenoptera Acutorostrata ◽  
Peptide Bonds ◽  
Pancreatic Ribonuclease

Pancreatic RNAase (ribonuclease) from the pike whale (lesser rorqual, Balaenoptera acutorostrata) was isolated by affinity chromatography. The protein was digested with different proteolytic enzymes. Peptides were isolated by gel filtration, preparative high-voltage paper electrophoresis and paper chromatography. The amino acid sequence of peptides was determined by the dansyl-Edman method. Although we do not have an amino acid composition for the whole protein, all peptide bonds were overlapped by one or more peptides. Residues 85-96 are bridged by a peptide of unstaisfactory composition and the sequence here depends, at least in part, on homology for its confirmation. Another region in which a similar situation obtains is residues 39-40. This pancreatic RNAase differs at 24-33% of the positions from all other mammalian pancreatic RNAases sequenced to date, except for pig RNAase, from which it differs by 19%. This indicates that whale RNAase has evolved independently during the larger part of the evolution of the mammals. Lesser-rorqual pancreatic RNAase is partially glycosidated (30%) at asparagine-76 in an Asn-Ser-Thr sequence (residues 76-78). Pig RNAase also has carbohydrate attached to asparagine-76 and is identical with lesser-rorqual RNAase in residues 76-98. Detailed evidence for the sequence has been deposited as Supplementary Publication SUP 50066 (11 pages) at the British Library Lending Division, Boston Spa, Wetherby, W. Yorkshire LS23 7BQ, U.K., from whom copies may be obtained on the terms ginen in Biochem. J. (1976) 135, 5.

Comparison of some properties of soil humic acids and synthetic phenolic polymers incorporating amino derivaties

Soil Research ◽  
1966 ◽  
Vol 4 (1) ◽  
pp. 41 ◽  
Author(s):  
JN Ladd ◽  
JHA Butler
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Acid Hydrolysis ◽  
Humic Acids ◽  
Gel Filtration ◽  
Quinone Imine ◽  
Peptide Bonds ◽  
Molecular Weights ◽  
Soil Humic Acids ◽  
Properties Of Soil

Twenty-three model phenolic polymers, either nitrogen-free or incorporating amino acids, peptides, or proteins, have been prepared from p-benzoquinone and catechol under mild oxidative conditions. Two lines of experimentation have demonstrated properties of soil humic acids closely similar to those of polymers incorporating proteins, but different from those of polymers incorporating amino acids: (1) fractionation of humic acids and synthetic polymers by 'Sephadex' gel filtration showed that the percentage of components of molecular weights nominally greater than 100 000 ranged from 52-76 % for eight humic acids tested, 53-59 % for benzoquinone-protein polymers (excluding polymers containing protamine), but less than 20% for all other polymers; (2) acid hydrolysis with 6M HCl resulted in a partial release of polymer nitrogen. Amino acid nitrogen in the hydrolysates accounted for 32.4-51.9 % of humic acid nitrogen, 31.2-56.3 % of the nitrogen of polymers incorporating protein, but less than 10.8% of the nitrogen of polymers incorporating individual amino acids. Experiments with model monomeric N-phenylglycine derivatives and with polymers incorporating simple peptides showed that the bond between the carbon atom of an aromatic ring and the nitrogen atom of an a-amino acid is far more stable to acid hydrolysis than peptide bonds or bonds linking amino acids in humic acids. Glycine is, however, readily released from N-phenylglycine derivatives when conditions favour their oxidation to a quinone-imine intermediate. Incorporation of proteins into phenolic polymers prevented the detection of peptide bonds by the Folin reagent.

scholarly journals The tryptic peptides of rabbit muscle triose phosphate isomerase

1974 ◽  
Vol 139 (1) ◽  
pp. 1-10 ◽  
Author(s):  
P. H. Corran ◽  
S. G. Waley
Keyword(s):  
Gel Filtration ◽  
Paper Electrophoresis ◽  
Triose Phosphate Isomerase ◽  
British Library ◽  
Rabbit Muscle ◽  
Amino Acid Residues ◽  
Tryptic Peptides ◽  
Triose Phosphate ◽  
Lending Library ◽  
Polypeptide Chains

1. The peptides obtained by tryptic digestion of S-[14C]carboxymethylated rabbit muscle triose phosphate isomerase have been studied. 2. The first step in the fractionation of the tryptic digest was gel filtration on coupled columns of Sephadex G-25 and G-50. Further fractionation was carried out by paper electrophoresis and paper chromatography. 3. The digest contained 26 peptides and three free amino acids. The sizes of the peptides ranged from two to 29 residues. 4. The sequences of the peptides have been determined. 5. The length of the polypeptide chains is about 250 amino acid residues. 6. The variant sequences encountered were due to partial deamidation; this may be one of the reasons for multiple forms of the enzyme. 7. The chicken and rabbit enzymes are compared. 8. Detailed evidence for the sequences of the tryptic peptides has been deposited as Supplementary Publication SUP 50024 at the British Library, Lending Division (formerly the National Lending Library for Science and Technology), Boston Spa, Yorks. LS23 7BQ, U.K., from whom copies can be obtained on the terms given in Biochem. J. (1973) 131, 5.

scholarly journals The Proteolytic Activity of Diapausing and Newly Hatched Red Mason Bees (Osmia Rufa: Megachilidae)

2014 ◽  
Vol 58 (1) ◽  
pp. 85-91
Author(s):  
Ewa A. Zaobidna ◽  
Kamila Dmochowska ◽  
Regina Frączek ◽  
Ewa Dymczyk ◽  
Krystyna Żółtowska
Keyword(s):  
Amino Acids ◽  
Enzyme Activity ◽  
Proteolytic Enzymes ◽  
Activity Levels ◽  
Solitary Bee ◽  
Peptide Bonds ◽  
Short Side ◽  
Male And Female ◽  
Substrate Preferences ◽  
Males And Females

Abstract Osmia rufa is a solitary bee that is used commercially for pollinating crops. The bee enters obligatory diapause as an imago. The activity of proteolytic enzymes during diapause has not been investigated. We studied the proteinase activity on four substrates - casein, haemoglobin, bovine serum albumin (BSA ), and gelatine - during diapause (from October to March) and in newly hatched males and females in April. During diapause, greater fluctuations in enzyme activity levels were noted in males than in females, and a significant decrease in male enzyme activity was observed in January and March. Male enzymes were most effective in decomposing gelatine; whereas, female enzymes were equally effective in hydrolysing gelatine and BSA . The differences in substrate preferences between male and female enzymes were particularly pronounced in October and in the newly hatched individuals. The levels of gelatinolytic activity likely indicate that a high proportion of proteinases in O. rufa are elastase-like enzymes. They are involved in the digestion and remodelling of proteins with numerous peptide bonds formed by amino acids with short side-chains.

scholarly journals Chemical and physical characterization of a proline-rich polypeptide from sheep colostrum

1981 ◽  
Vol 199 (1) ◽  
pp. 9-15 ◽  
Author(s):  
M Janusz ◽  
K Starościk ◽  
M Zimecki ◽  
Z Wieczorek ◽  
J Lisowski
Keyword(s):  
Amino Acids ◽  
Molecular Weight ◽  
Room Temperature ◽  
Proteolytic Enzymes ◽  
Gel Filtration ◽  
Guanidinium Chloride ◽  
Terminal Amino Acid ◽  
Polyproline Ii ◽  
Terminal Amino

A proline-rich polypeptide isolated from sheep colostrum is described. The molecular weight of the polypeptide determined by gel filtration is 17 200. However, in the presence of guanidinium chloride the molecular weight found is about 6000. The polypeptide contains about 22% of proline, a high proportion of non-polar amino acids, a low percentage of glycine, and no alanine, arginine and cysteine residues. The only N-terminal amino acid found is leucine. C.d. spectra in water and in 50% (v/v) trifluoroethanol suggest the presence of block sequences of proline residues forming helices of polyproline II type. The proline-rich polypeptide is soluble at 4 degrees C but is reversibly precipitated on warming to room temperature. Maximal precipitation is observed at pH 4.6 and at ionic strength above 0.6. The precipitation depends on the concentration of the polypeptide. No effect of other proteins, Ca2+ and Zn2+ ions on the precipitation of the polypeptide was found. The proline-rich polypeptide is not an amphipathic protein. The lack of effect of the polypeptide on proteolytic enzymes ruled out the possibility that it is an inhibitor of proteinases.

scholarly journals Some properties of the lectin from Datura stramonium (thorn-apple) and the nature of its glycoprotein linkages

1981 ◽  
Vol 197 (2) ◽  
pp. 345-353 ◽  
Author(s):  
Nila N. Desai ◽  
Anthony K. Allen ◽  
Albert Neuberger
Keyword(s):  
Amino Acids ◽  
Polypeptide Chain ◽  
Gel Filtration ◽  
Datura Stramonium ◽  
Inhibitory Effect ◽  
Plant Cell Walls ◽  
Thorn Apple ◽  
A Subunit ◽  
Sepharose 4B ◽  
Derivatives Of

The lectin from Datura stramonium (thorn-apple; Solanaceae) has been purified by affinity chromatography and shown to be a glycoprotein containing about 40% (w/w) of carbohydrate. The most abundant amino acids are hydroxyproline, cystine, glycine and serine. Results obtained by gel filtration in 6m-guanidinium chloride on Sepharose 4B suggest that it has a subunit mol.wt. of about 30000 and that it probably associates into dimers. The lectin is inhibited specifically by chitin oligosaccharides and bacterial-cell-wall oligosaccharides, but only weakly by N-acetylglucosamine. Glycopeptides from soya-bean (Glycine max) lectin and fetuin are also strong inhibitors of Datura lectin, indicating that it interacts with internal N-acetylglucosamine residues. Its specificity is similar to, but not identical with, that of potato (Solanum tuberosum) lectin. After prolonged proteolytic digestion of reduced and S-carboxymethylated or S-aminoethylated derivatives of the lectin, glycopeptides of mol.wt. of about 18000 were isolated. The glycopeptides contained all the carbohydrate and hydroxyproline of the original glycoprotein, and lesser amounts of serine, S-carboxymethylcysteine and other amino acids. The arabinose residues of the glycoprotein are present as β-l-arabinofuranosides linked to the polypeptide chain through the hydroxyproline residues, and can be removed by mild acid treatment; the ratio of arabinose to hydroxyproline is 3.4:1. Some of the serine residues of the polypeptide chain are substituted with one or two α-galactopyranoside residues, most of which can be removed by the action of α-galactosidase. The galactose residues are more easily removed from the acid-treated glycopeptide (from which arabinose has been removed) than from the complete glycopeptide, indicating a steric hindrance of the galactosidase action by the adjacent chains of arabinosides. There is a slow release of galactose residues by a process of β-elimination in 0.5m-NaOH (pH13.7) from the complete glycopeptide, and a fairly rapid release of galactose by this process from the acid-treated glycopeptide, which lacks arabinose. This is probably due to the inhibitory effect of the negative charge on the adjacent arabinofuranoside residues. The similarities and differences between the lectins from Datura and potato are discussed, as are their structural resemblance to glycopeptides that have been isolated from plant cell walls.

Release of amino acids from soil humic acids by proteolytic enzymes

Soil Research ◽  
1967 ◽  
Vol 5 (2) ◽  
pp. 161 ◽  
Author(s):  
JN Ladd ◽  
PG Brisbane
Keyword(s):  
Amino Acids ◽  
Acid Hydrolysis ◽  
Humic Acids ◽  
Direct Evidence ◽  
Proteolytic Enzymes ◽  
Approximate Inverse ◽  
Peptide Bonds ◽  
Aromatic Content ◽  
Phenolic Polymer ◽  
Soil Humic Acids

Direct evidence for the occurrence of peptide bonds in soil humic acids has been obtained by showing that the proteolytic enzyme pronase releases a-amino acids from each of 10 humic acids. The maximal amounts of amino acids liberated by pronase accounted for 27.6-39.4% of those released by hot acid hydrolysis. The susceptibility of humic acids to proteolytic attack showed an approximate inverse relationship with their aromatic content, based on their absorbance at 260 or 450 m�. Both pronase and papain attacked casein less readily when it was incorporated into a phenolic polymer prepared from p-benzoquinone. Papain was inactive towards soil humic acids. Similar amino acids were released by pronase from each of five humic acids tested. The amino acid patterns showed no basic amino acids but relatively high proportions of leucine (isoleucine) as compared with those obtained by acid hydrolysis. The action of pronase on humic acids had little effect on the molecular weight distribution of the humic components.

Proteolysis in Cheddar cheese: role of coagulant and starter bacteria

1978 ◽  
Vol 45 (3) ◽  
pp. 465-477 ◽  
Author(s):  
Arthur M. O'Keeffe ◽  
Patrick F. Fox ◽  
Charles Daly
Keyword(s):  
Amino Acids ◽  
High Voltage ◽  
Free Amino Acids ◽  
Free Amino ◽  
Protein Solubility ◽  
Gel Filtration ◽  
Cheddar Cheese ◽  
Paper Electrophoresis ◽  
Small Peptides

SummaryCheddar cheese was produced free of non-starter bacteria, acidified with starter or glucono-δ-lactone and containing active coagulant (chymosin or pepsin) or inactivated coagulant (pepsin). The level and type of proteolysis in the experimental cheeses was monitored by protein solubility at pH 4·6 and in 12 % TCA, polyacrylamide gel and high voltage paper electrophoresis, gel filtration and paper chromatography. The results show that the coagulant was primarily responsible for the formation of large peptides while small peptides and free amino acids were produced principally by the starter, possibly from coagulant-produced peptides.

Digestive processes of haematophagous insects. XIII. Evidence for the digestive function of midgut proteinases of Glossina morsitans morsitans Westwood (Diptera: Glossinidae)

1977 ◽  
Vol 55 (9) ◽  
pp. 1557-1562 ◽  
Author(s):  
R. H. Gooding
Keyword(s):  
Amino Acids ◽  
Free Amino Acids ◽  
Free Amino ◽  
Proteolytic Enzymes ◽  
Gel Filtration ◽  
Glossina Morsitans Morsitans ◽  
Glossina Morsitans ◽  
Anterior Section ◽  
Digestive Function ◽  
Posterior Section

Trypsin (EC 3.4.21.4), proteinase VI, proteinase VII, aminopeptidase (EC 3.4.11.1), carboxypeptidase A (EC 3.4.12.2), and carboxypeptidase B (EC 3.4.11.3) occur in the posterior section of the midgut of unfed male and female Glossina morsitans morsitans Westwood, and their activities rise after a blood meal. Only traces of these enzymes occur in the anterior section of the midgut. Elution profiles of proteins during Sephadex gel filtration of the anterior midgut at various times after feeding, and the low ratio of free amino acids to protein in the lumen of the anterior section of the midgut indicate that no significant hydrolysis of protein takes place there. Results of Sephadex® gel filtration of material from the posterior section of the midgut indicate that proteins are rapidly converted to peptides and free amino acids, which occur in high concentration there. The results are interpreted as indicating that digestion of proteins takes place only in the posterior section of the midgut, and that all six proteolytic enzymes have a digestive function.

scholarly journals Isolation and characterization of a hydroxyproline-containing protein from soluble extracts of the leaves of sandal (Santalum album L.)

1974 ◽  
Vol 141 (1) ◽  
pp. 147-153 ◽  
Author(s):  
Uliyar V. Mani ◽  
Amurtur N. Radhakrishnan
Keyword(s):  
Amino Acids ◽  
Proteolytic Enzymes ◽  
Gel Filtration ◽  
Disc Electrophoresis ◽  
Partial Hydrolysis ◽  
Santalum Album ◽  
Isolation And Characterization ◽  
Approximate Molecular Weight ◽  
Santalum Album L

1. A hydroxyproline-containing protein was isolated from the soluble fraction of sandal leaves (Santalum album L.) and the purified protein was homogeneous by disc electrophoresis. 2. It is a glycoprotein containing 16% carbohydrate, the components of which were mainly arabinose, with only small amounts (about 5%) of galactose. The principal amino acids were glutamic acid, aspartic acid, glycine, alanine, arginine, lysine, proline and hydroxyproline, which together comprised 60% of the total. The number of acidic amino acids exceeds the number of basic amino acids. By Sephadex gel filtration, the approximate molecular weight was found to be about 63000. The ratio of residues of hydroxyproline to those of arabinose was 1:2. 3. The native protein is resistant to the action of several proteolytic enzymes. After partial hydrolysis with 0.1m-HCl, the protein became susceptible to attack by Pronase but remained resistant to collagenase.

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