The Amino Acid Sequence of Streptomyces griseus Protease A: The Peptic Peptides

The peptic peptides of Streptomyces griseus Protease A (excluding the previously characterized disulfide bridge peptic peptides) were fractionated into basic, neutral, and neutral plus acidic peptide fractions by chromatography on Dowex 1 × 2. These three peptide fractions were then fractionated by cation-exchange chromatography on Chromobead P resin using the Technicon autoanalyzer system. Following further purifications on paper, the amino acid compositions and sequences of the peptic peptides were determined. The N-terminal sequence of Protease A has been identified as Ile–Ala–Gly–Gly–Glu–Ala. The numbers of amino acid residues obtained from the amino acid sequences reported are in agreement with those numbers obtained from amino acid analysis of the total protein in the cases of tryptophan, methionine, histidine, proline, phenylalanine, and glutamic acid. Some of the results suggest either the presence of nonidentical but highly homologous proteins in the Protease A preparation or the possibility of repeating sequences in a single molecular species.

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A NEW PEPTIDE THROMBIN INHIBITOR FROM STREPTOMYCES GRISEUS

10.1055/s-0038-1644330 ◽

1987 ◽

Author(s):

Hua-zhen Liu ◽

Wei Chen ◽

Qi-ying Liu ◽

Xia Zhang ◽

Li-xiu Wang ◽

...

Keyword(s):

Amino Acid ◽

High Performance ◽

Antithrombin Iii ◽

Streptomyces Griseus ◽

Exchange Chromatography ◽

Performic Acid ◽

Thrombin Inhibitor ◽

Amino Acid Residues ◽

Macroporous Resin ◽

Terminal Residue

A new peptide thrombin inhibitor was found in the Streptomyces griseus strain 254 isolated from a soil sample from Tongan, Fujian province, China, the inhibitor being a secondary metabolic product. The production of the inhibitor reached a maximum after 3 days culture of bacteria at 28°C in a rotary shaker. The inhibitor excreted in the culture filtrate was purified by absorption on macroporous resin, followed by ion exchange chromatography on DEAE-52, CM-32 cellulose, affinity chromatography on the immobilized thrombin and high performance liquid chromatography. The amino acid composition of the inhibitor was determined to be Val(2), Met(l), Ile(l), Leu(2) and Arg (1), similar to that of the amino acid residues around the reactive site of human antithrombin III, the critical plasma inhibitor of thrombin. The NH2-terminal residue of the inhibitor seems to be blocked by the alkyl group due to the negative reaction to ninhydrin, whereas the COO-terminal residue is most likely to be arginal because of that Arg was not found in the amino acid analysis, unless the peptide was oxidized by performic acid before acid hydrolysis. The chromogen substrates Bz-Phe-Val-Arg-PNA and Bz-Gly-Pro-Lys-PNA were used to determine the thrombin and plasmin activities, respectively. Besides thrombin, the purified inhibitor also exhibits a weak inhibitory activities on trypsin and much weak on plasmin, but not on chymotrypsin and other protein-ases.

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Isolation of Two Bovine Amelogenin Peptides and Their Amino Acid Sequences

Advances in Dental Research ◽

10.1177/08959374870010021701 ◽

1987 ◽

Vol 1 (2) ◽

pp. 276-281 ◽

Cited By ~ 5

Author(s):

J.-H. Yeh ◽

T. Takagi ◽

S. Sasaki

Keyword(s):

Molecular Weight ◽

Amino Acid ◽

Gel Electrophoresis ◽

Polyacrylamide Gel Electrophoresis ◽

Amino Acid Sequences ◽

Polyacrylamide Gel ◽

The Other ◽

Edman Degradation ◽

Amino Acid Residues ◽

Peptide Fractions

Two peptide fractions of bovine amelogenin having a highly aggregative property to form polymers were purified by chromatography, SDS-polyacrylamide gel electrophoresis, and HPLC. Amino acid sequences of purified peptides were determined by automated Edman degradation. One peptide was found to be composed of 63 amino acid residues having a molecular weight of 7105, and the other of 86 residues having that of 9683. The sequence of the smaller peptide was identical to the C-terminal 63 residues of the amelogenin molecule of 170 residues previously reported, but the larger contained eight residues which are absent in the amelogenin sequence. There is a possibility that the latter peptide might be synthesized independently from mRNA spliced at different positions.

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Two globin strains in the giant annelid extracellular haemoglobins

Biochemical Journal ◽

10.1042/bj2410441 ◽

1987 ◽

Vol 241 (2) ◽

pp. 441-445 ◽

Cited By ~ 35

Author(s):

T Gotoh ◽

F Shishikura ◽

J W Snow ◽

K I Ereifej ◽

S N Vinogradov ◽

...

Keyword(s):

Amino Acid ◽

Lumbricus Terrestris ◽

Ion Exchange Chromatography ◽

Exchange Chromatography ◽

Amino Acid Sequences ◽

Amino Acid Residues ◽

Terminal Amino Acid ◽

Reverse Phase ◽

Terminal Amino ◽

Polypeptide Chains

The constituent polypeptide chains I, II, III and IV of the giant extracellular haemoglobin of the oligochaete Lumbricus terrestris were isolated by mono Q ion-exchange chromatography and C8 reverse-phase chromatography. The N-terminal amino acid sequences of Lumbricus chains I, III and IV were determined and aligned with those of Lumbricus chain II and the four chains of the extracellular haemoglobin of the polychaete Tylorrhynchus heterochaetus. Three invariant amino acid residues, Cys-7, Val-15 and Trp-19, were found to occur in the N-terminal segments (17-22 residues) of the eight chains of Lumbricus and Tylorrhynchus haemoglobins. In addition, it was found that the eight sequences could be separated into two groups: ‘A’, consisting of Lumbricus chains I and II and Tylorrhynchus chains I and IIA, having invariant Lys-14 and Lys-16, and ‘B’, consisting of Lumbricus chains III and IV and Tylorrhynchus IIB and IIC, having invariant Cys-6, Ser-8 and Asp-11. This result suggests that there are two strains of globin chain in the annelid extracellular haemoglobins.

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The Amino Acid Sequence of Streptomyces griseus Trypsin. I. The Peptic Peptides

Canadian Journal of Biochemistry ◽

10.1139/o73-141 ◽

1973 ◽

Vol 51 (7) ◽

pp. 1077-1088 ◽

Cited By ~ 1

Author(s):

L. Jurášek ◽

L. B. Smillie

Keyword(s):

Amino Acid ◽

Serine Proteases ◽

System Analysis ◽

Streptomyces Griseus ◽

Ion Exchange Chromatography ◽

Exchange Chromatography ◽

Amino Acid Sequences ◽

Terminal Sequence ◽

Unique Amino Acid ◽

High Voltage Electrophoresis

Streptomyces griseus trypsin (S.G.T.) isolated from pronase was digested with pepsin. The peptic peptides were isolated by high-voltage electrophoresis on paper and ion-exchange chromatography on Chromobead P resin using the Technicon autoanalyzer system. Analysis of the purified peptides provides 28 unique amino acid sequences accounting for approximately 95% of the S.G.T. molecule. A portion of the residues not accounted for can be ascribed to free leucine, phenylalanine, tyrosine, and tryptophan present in the peptic digest. The NH2-terminal sequence of S.G.T. was shown to be Val–Val–Gly–Gly–Thr–Arg–Ala–Ala–Gln–Gly–Glu–Phe and is highly homologous with NH2-terminal sequences of other Asp–Ser–Gly serine proteases.

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Amino acid sequences of peptides from a chymotryptic digest of a urea-soluble protein fraction (U.S.3) from oxidized wool

Biochemical Journal ◽

10.1042/bj1150323 ◽

1969 ◽

Vol 115 (2) ◽

pp. 323-334 ◽

Cited By ~ 6

Author(s):

M. C. Corfield ◽

J. C. Fletcher

Keyword(s):

Amino Acid ◽

Cation Exchange ◽

Protein Fraction ◽

Exchange Resin ◽

Cation Exchange Resin ◽

Paper Electrophoresis ◽

Amino Acid Sequences ◽

Amino Acid Residues ◽

Soluble Protein Fraction ◽

Peptide Fractions

1. A chymotryptic digest of the protein fraction U.S.3. from oxidized wool was separated into 51 peptide fractions by chromatography on a column of cation-exchange resin. 2. The less acidic fractions were separated into their component peptides by a combination of cation-exchange-resin chromatography, paper chromatography and paper electrophoresis. 3. The amino acid sequences of 34 of these peptides were elucidated, and those of 14 others partially determined. 4. Overlaps between the tryptic and chymotryptic peptides from fraction U.S.3 have enabled ten extended amino acid sequences to be deduced, the longest containing 20 amino acid residues. 5. The relevance of the results to the structures of the helical and non-helical regions of wool is discussed.

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Identification and Characterization of Lactocyclicin Q, a Novel Cyclic Bacteriocin Produced by Lactococcus sp. Strain QU 12

Applied and Environmental Microbiology ◽

10.1128/aem.02299-08 ◽

2009 ◽

Vol 75 (6) ◽

pp. 1552-1558 ◽

Cited By ~ 81

Author(s):

Naruhiko Sawa ◽

Takeshi Zendo ◽

Junko Kiyofuji ◽

Koji Fujita ◽

Kohei Himeno ◽

...

Keyword(s):

Amino Acid ◽

High Performance ◽

Hydrophobic Interaction Chromatography ◽

Heat Stability ◽

Peptide Bond ◽

Exchange Chromatography ◽

Amino Acid Sequences ◽

Amino Acid Residues ◽

Gene Encoding ◽

High Level

ABSTRACT Lactococcus sp. strain QU 12, which was isolated from cheese, produced a novel cyclic bacteriocin termed lactocyclicin Q. By using cation-exchange chromatography, hydrophobic interaction chromatography, and reverse-phase high-performance liquid chromatography, lactocyclicin Q was purified from culture supernatant, and its molecular mass was determined to be 6,062.8 Da by mass spectrometry. Lactocyclicin Q has been characterized by its unique antimicrobial spectrum, high level of protease resistance, and heat stability compared to other reported bacteriocins of lactic acid bacteria. The amino acid sequence of lactocyclicin Q was determined chemically, and this compound is composed of 61 amino acid residues that have a cyclic structure with linkage between the N and C termini by a peptide bond. It showed no homology to any other antimicrobial peptide, including cyclic bacteriocins. On the basis of the amino acid sequences obtained, the sequence of the gene encoding the prepeptide lactocyclicin Q was obtained. This is the first report of a cyclic bacteriocin purified from a strain belonging to the genus Lactococcus.

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A spatial similarity of stereochemical environments formed by amino acid residues defines a common epitope of two non-homologous proteins

Scientific Reports ◽

10.1038/s41598-019-51350-2 ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 1

Author(s):

Kentaro Nakashima ◽

Shintaro Iwashita ◽

Takehiro Suzuki ◽

Chieko Kato ◽

Toshiyuki Kohno ◽

...

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Cultured Cells ◽

Amino Acid Sequences ◽

Cross Reaction ◽

Amino Acid Residues ◽

Homologous Proteins ◽

Spatial Similarity ◽

Target Molecules ◽

The Cross

Abstract It is critical for development of high-quality antibodies in research and diagnostics to predict accurately their cross-reactivities with “off-target” molecules, which potentially induce false results. Herein, we report a good example of such a cross-reactivity for an off-target due to a stereochemical environment of epitopes, which does not simply depend on amino acid sequences. We found that significant subpopulation of a polyclonal peptide antibody against Bcnt (Bucentaur) (anti-BCNT-C antibody) cross-reacted with a completely different protein, glutamine synthetase (GS), and identified four amino acids, GYFE, in its C-terminal region as the core amino acids for the cross-reaction. Consistent with this finding, the anti-BCNT-C antibody strongly recognized endogenously and exogenously expressed GS in tissues and cultured cells by Western blotting and immunohistochemistry. Furthermore, we elucidated that the cross-reaction is caused by a spatial similarity between the stereochemical environments formed by amino acid residues, including the GYFE of GS and the GYIE of Bcnt, rather than by their primary sequences. These results suggest it is critical to comprehensively analyze antibody interactions with target molecules including off-targets with special attention to the physicochemical environments of epitope-paratope interfaces to decrease the risk of false interpretations of results using antibodies in science and clinical applications.

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Structural Relationship Between Human Salivary Histatins

Journal of Dental Research ◽

10.1177/00220345900690010101 ◽

1990 ◽

Vol 69 (1) ◽

pp. 2-6 ◽

Cited By ~ 120

Author(s):

R.F. Troxler ◽

G.D. Offner ◽

T. Xu ◽

J.C. Vanderspek ◽

F.G. Oppenheim

Keyword(s):

Amino Acid ◽

Amino Acid Sequences ◽

Amino Acid Residues ◽

Homologous Proteins ◽

Pathogenic Yeast ◽

Histatin 5 ◽

Structural Relationships ◽

Carboxyl Terminal ◽

Relationship Of ◽

First Time

Histatins are a group of electrophoretically distinct histidine-rich polypeptides with microbicidal activity found in human parotid and submandibular gland secretions. Recently, we have shown that histatins 1, 3, and 5 are homologous proteins that consist of 38, 32, and 24 amino acid residues, respectively, and that these polypeptides kill the pathogenic yeast, Candida albicans. We now describe the isolation and structural characterization of histatins 2, 4, 6, and 7-12, the remaining members of this group of polypeptides. Histatin 2 was found to be identical to the carboxyl terminal 26 residues of histatin 1; histatin 4 was found to be identical to the carboxyl terminal 20 residues of histatin 3; and histatin 6 was found to be identical to histatin 5, but contained an additional carboxyl terminal arginine residue. The amino acid sequences of histatins 7-12 formally correspond to residues 12-24, 13-24, 12-25, 13-25, 5-11, and 5-12, respectively, of histatin 3, but could also arise proteolytically from histatin 5 or 6. These results establish, for the first time, the complete structural relationships between all members of this group of microbicidal proteins in human parotid saliva. The relationship of histatins to one another is discussed in the context of their genetic origin, biosynthesis and secretion into the oral cavity, and potential as reagents in anti-candidal studies.

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Bovine Factors X1And X2: Activation Peptide Based Chromatographic Differences

10.1055/s-0038-1665665 ◽

1979 ◽

Author(s):

Takashi Morita ◽

Craig Jackson

Keyword(s):

Amino Acid ◽

Anion Exchange ◽

Gel Filtration ◽

Heart Association ◽

Personal Communication ◽

Exchange Chromatography ◽

Amino Acid Sequences ◽

Factor X ◽

Activation Peptide ◽

Activation Peptides

Bovine Factor X is eluted in two forms (X1and X2) from anion exchange chromatographic columns. These two forms have indistinguishable amino acid compositions, molecular weights and specific activities. The amino acid sequences containing the γ-carboxyglutamic acid residues have been shown to be identical in X1 and X2(H. Morris, personal communication). An activation peptide is released from the N-terminal region of the heavy chain of Factor X by an activator from Russell’s viper venom. This peptide can be isolated after activation by gel filtration on Sephadex G-100 under nondenaturing conditions. The activation peptides from a mixture of Factors X1 and X2 were separated into two forms by anion-exchange chromatography. The activation peptide (AP1) which eluted first was shown to be derived from Factor X1. while the activation peptiae (AP2) which eluted second was shown to be derived from X2 on the basis of chromatographic separations carried out on Factors X1 and X2 separately. Factor Xa was eluted as a symmetrical single peak. On the basis of these and other data characterizing these products, we conclude that the difference between X1 and X2 are properties of the structures of the activation peptides. (Supported by a grant HL 12820 from the National Heart, Lung and Blood Institute. C.M.J. is an Established Investigator of the American Heart Association).

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Computational Analysis of Therapeutic Enzyme Uricase from Different Source Organisms

Current Proteomics ◽

10.2174/1570164616666190617165107 ◽

2020 ◽

Vol 17 (1) ◽

pp. 59-77

Author(s):

Anand Kumar Nelapati ◽

JagadeeshBabu PonnanEttiyappan

Keyword(s):

Uric Acid ◽

Amino Acid ◽

Sequence Alignment ◽

Multiple Sequence Alignment ◽

Protein Sequences ◽

Amino Acid Sequences ◽

Amino Acid Residues ◽

Multiple Sequence ◽

Physiochemical Properties ◽

Pharmaceutical Industries

Background:Hyperuricemia and gout are the conditions, which is a response of accumulation of uric acid in the blood and urine. Uric acid is the product of purine metabolic pathway in humans. Uricase is a therapeutic enzyme that can enzymatically reduces the concentration of uric acid in serum and urine into more a soluble allantoin. Uricases are widely available in several sources like bacteria, fungi, yeast, plants and animals.Objective:The present study is aimed at elucidating the structure and physiochemical properties of uricase by insilico analysis.Methods:A total number of sixty amino acid sequences of uricase belongs to different sources were obtained from NCBI and different analysis like Multiple Sequence Alignment (MSA), homology search, phylogenetic relation, motif search, domain architecture and physiochemical properties including pI, EC, Ai, Ii, and were performed.Results:Multiple sequence alignment of all the selected protein sequences has exhibited distinct difference between bacterial, fungal, plant and animal sources based on the position-specific existence of conserved amino acid residues. The maximum homology of all the selected protein sequences is between 51-388. In singular category, homology is between 16-337 for bacterial uricase, 14-339 for fungal uricase, 12-317 for plants uricase, and 37-361 for animals uricase. The phylogenetic tree constructed based on the amino acid sequences disclosed clusters indicating that uricase is from different source. The physiochemical features revealed that the uricase amino acid residues are in between 300- 338 with a molecular weight as 33-39kDa and theoretical pI ranging from 4.95-8.88. The amino acid composition results showed that valine amino acid has a high average frequency of 8.79 percentage compared to different amino acids in all analyzed species.Conclusion:In the area of bioinformatics field, this work might be informative and a stepping-stone to other researchers to get an idea about the physicochemical features, evolutionary history and structural motifs of uricase that can be widely used in biotechnological and pharmaceutical industries. Therefore, the proposed in silico analysis can be considered for protein engineering work, as well as for gout therapy.

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