Macedocin, a Food-Grade Lantibiotic Produced by Streptococcus macedonicus ACA-DC 198

ABSTRACT Streptococcus macedonicus ACA-DC 198, a strain isolated from Greek Kasseri cheese, produces a food-grade lantibiotic named macedocin. Macedocin has a molecular mass of 2,794.76 ± 0.42 Da, as determined by electrospray mass spectrometry. Partial N-terminal sequence analysis revealed 22 amino acid residues that correspond with the amino acid sequence of the lantibiotics SA-FF22 and SA-M49, both of which were isolated from the pathogen Streptococcus pyogenes. Macedocin inhibits a broad spectrum of lactic acid bacteria, as well as several food spoilage and pathogenic bacteria, including Clostridium tyrobutyricum. It displays a bactericidal effect towards the most sensitive indicator strain, Lactobacillus sakei subsp. sakei LMG 13558T, while the producer strain itself displays autoinhibition when it is grown under conditions that do not favor bacteriocin production. Macedocin is active at pHs between 4.0 and 9.0, and it retains activity even after incubation for 20 min at 121°C with 1 atm of overpressure. Inhibition of macedocin by proteolytic enzymes is variable.

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Damage of Amino Acid Residues of Proteins after Reaction with Oxidizing Lipids: Estimation by Proteolytic Enzymes

Journal of Food Science ◽

10.1111/j.1365-2621.1984.tb10397.x ◽

1984 ◽

Vol 49 (4) ◽

pp. 1082-1084 ◽

Cited By ~ 9

Author(s):

TERUYOSHI MATOBA ◽

DAIZO YONEZAWA ◽

BABOO M. NAIR ◽

MAKOTO KITO

Keyword(s):

Amino Acid ◽

Proteolytic Enzymes ◽

Amino Acid Residues

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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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The Primary Structure of Antithrombin-III

10.1055/s-0039-1680621 ◽

1977 ◽

Author(s):

T. E. Petersen ◽

G. Dudek-Wojciechowska ◽

L. Sottrup-Jensen ◽

S. Magnusson

Keyword(s):

Amino Acid ◽

Antithrombin Iii ◽

Factor Xa ◽

Amino Acid Residues ◽

Disulfide Bridges ◽

Single Chain ◽

Terminal Sequence ◽

Chymotryptic Peptide ◽

Sequence Region ◽

Bovine Factor

Human antithrombin-III is a single-chain glycoprotein with three disulfide bridges and four prosthetic glucosamine-based oligosaccharide groups. The disulfide bridges have been established. In four fragments of 208, 168, 3 and 46 amino acid residues, resp. 415 of the appr. 425 residues have been sequenced. The four oligosaccharide groups are attached to four Asn-residues within a sequence region of 95 residues. No extensive sequence homology with the trypsin inhibitors has been observed. One chymotryptic peptide was found to be a substrate for bovine factor Xa, cleaving the arginyl bond in the sequence -Ile-Val-Ala-Glu-Gly-Arg-Asp-. A second peptide is cleaved by thrombin. It is not clear whether these sites are inhibitor sites in the native molecule. Other possible candidates for inhibitor sites are a -Val-Leu-Ile-Leu-Pro-Lys-Pro- sequence (similar to the sequence 40-48 of hirudin, which also includes a -Pro-Lys-Pro- sequence) and also the C-terminal sequence -Gly-Arg-Val-Ala-Asn-Pro-Cys-Val-Lys.

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Hyaluronan-binding region of aggrecan from pig laryngeal cartilage. Amino acid sequence, analysis of N-linked oligosaccharides and location of the keratan sulphate

Biochemical Journal ◽

10.1042/bj2860761 ◽

1992 ◽

Vol 286 (3) ◽

pp. 761-769 ◽

Cited By ~ 31

Author(s):

F P Barry ◽

J U Gaw ◽

C N Young ◽

P J Neame

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Amino Acid Sequence ◽

Signal Peptidase ◽

Amino Acid Residues ◽

Terminal Sequence ◽

Binding Region ◽

Keratan Sulphate ◽

Laryngeal Cartilage ◽

Hyaluronan Binding

The hyaluronan-binding region (HABR) was prepared from pig laryngeal cartilage aggrecan and the amino acid sequence was determined. The HABR had two N-termini: one N-terminal sequence was Val-Glu-Val-Ser-Glu-Pro (367 amino acids in total), and a second N-terminal sequence (Ala-Ile-Ser-Val-Glu-Val; 370 amino acids in total) was found to arise due to alternate cleavage by the signal peptidase. The N-linked oligosaccharides were analysed by examining their reactivity with a series of lectins. It was found that the N-linked oligosaccharide on loop A was of the mannose type, while that on loop B was of the complex type. No reactivity was detected between the N-linked oligosaccharide on loop B' and any of the lectins. The location of keratan sulphate (KS) in the HABR was determined by Edman degradation of the immobilized KS-containing peptide. The released amino acid derivatives were collected and tested for the presence of epitope to antibody 5-D-4. On the basis of 5-D-4 reactivity and sequencing yields, the KS chains are attached to threonine residues 352 and 357. There is no KS at threonine-355. This site is not in fact in G1, but about 16 amino acid residues into the interglobular domain. Comparison of the structure of the KS chain from the HABR and from the KS domain of pig laryngeal cartilage aggrecan was made by separation on polyacrylamide gels of the oligosaccharides arising from digestion with keratanase. Comparison of the oligosaccharide maps suggests that the KS chains from both parts of the aggrecan molecule have the same structure.

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Septal Pore Cap Protein SPC18, Isolated from the Basidiomycetous Fungus Rhizoctonia solani, Also Resides in Pore Plugs

Eukaryotic Cell ◽

10.1128/ec.00125-08 ◽

2008 ◽

Vol 7 (10) ◽

pp. 1865-1873 ◽

Cited By ~ 21

Author(s):

Kenneth G. A. van Driel ◽

Arend F. van Peer ◽

Jan Grijpstra ◽

Han A. B. Wösten ◽

Arie J. Verkleij ◽

...

Keyword(s):

Endoplasmic Reticulum ◽

Amino Acid ◽

Rhizoctonia Solani ◽

Filamentous Fungi ◽

Signal Peptide ◽

Biochemical Composition ◽

Amino Acid Residues ◽

Terminal Sequence ◽

Basidiomycetous Fungus ◽

Central Pore

ABSTRACT The hyphae of filamentous fungi are compartmentalized by septa that have a central pore. The fungal septa and septum-associated structures play an important role in maintaining cellular and intrahyphal homeostasis. The dolipore septa in the higher Basidiomycota (i.e., Agaricomycotina) are associated with septal pore caps. Although the ultrastructure of the septal pore caps has been studied extensively, neither the biochemical composition nor the function of these organelles is known. Here, we report the identification of the glycoprotein SPC18 that was found in the septal pore cap-enriched fraction of the basidiomycetous fungus Rhizoctonia solani. Based on its N-terminal sequence, the SPC18 gene was isolated. SPC18 encodes a protein of 158 amino acid residues, which contains a hydrophobic signal peptide for targeting to the endoplasmic reticulum and has an N-glycosylation motif. Immunolocalization showed that SPC18 is present in the septal pore caps. Surprisingly, we also observed SPC18 being localized in some plugs. The data reported here strongly support the hypothesis that septal pore caps are derived from endoplasmic reticulum and are involved in dolipore plugging and, thus, contribute to hyphal homeostasis in basidiomycetous fungi.

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Characterization of the sulfated glycopeptide of chicken pepsinogen

Collection of Czechoslovak Chemical Communications ◽

10.1135/cccc19820709 ◽

1982 ◽

Vol 47 (2) ◽

pp. 709-718 ◽

Cited By ~ 6

Author(s):

Miroslav Baudyš ◽

Vladimír Kostka ◽

Karel Grüner ◽

Jan Pohl

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Protein Molecule ◽

Amino Acid Residues ◽

Terminal Sequence ◽

Sugar Moiety ◽

Pepsinogen A ◽

Identical Amino Acid Sequence ◽

High Voltage Electrophoresis

S-sulfonated chicken pepsinogen was digested with TPCK-trypsin; large tryptic peptides, separated on Sephadex G-25 fine, were subjected to additional cleavage with α-chymotrypsin. The hold-up fraction of the chymotryptic digest from the Sephadex G-25 column, was resolved by high voltage electrophoresis. The three most acidic zones contained glycopeptides of identical amino acid sequence Val-Ser-Thr-Asn-Glu-Thr-Val-Tyr, yet differed in the composition of the sugar moiety. These glycopeptides, moreover, bear different numbers of sulfate groups which enabled the resolution of the peptides. The most acidic glycopeptide contains 7 glucosamine residues, 3 mannose residues and 5 sulfate groups, the second one 6 glucosamine residues, 3 mannose residues and 4 sulfate groups and the slowest, minority glycopeptide, 5 glucosamine residues, 2 mannose residues and 2 sulfate groups. The entire sugar moiety is attached to one of the chain viaasparagine. In other experiments the glycopeptides were also isolated from the thermolytic digest of chicken pepsin; their C-terminal sequence was shorter by two amino acid residues. The tentative assignment of the glycopeptides to the amino acid sequence of pepsinogen resulted from the analysis of the limited tryptic digest of the whole protein molecule. Chicken pepsinogen is glycosylated at the site of the chain occupied by a phosphoserine residue in hog pepsinogen A.

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Purification, characterization and partial amino acid sequence of glycogen synthase from Saccharomyces cerevisiae

Biochemical Journal ◽

10.1042/bj2680401 ◽

1990 ◽

Vol 268 (2) ◽

pp. 401-407 ◽

Cited By ~ 9

Author(s):

A Carabaza ◽

J Arino ◽

J W Fox ◽

C Villar-Palasi ◽

J J Guinovart

Keyword(s):

Saccharomyces Cerevisiae ◽

Amino Acid ◽

Glycogen Synthase ◽

Amino Acid Residues ◽

Terminal Sequence ◽

Muscle Enzyme ◽

Western Blots ◽

Enzyme Sequence ◽

A Subunit ◽

Yeast Glycogen

Glycogen synthase from Saccharomyces cerevisiae was purified to homogeneity. The enzyme showed a subunit molecular mass of 80 kDa. The holoenzyme appears to be a tetramer. Antibodies developed against purified yeast glycogen synthase inactivated the enzyme in yeast extracts and allowed the detection of the protein in Western blots. Amino acid analysis showed that the enzyme is very rich in glutamate and/or glutamine residues. The N-terminal sequence (11 amino acid residues) was determined. In addition, selected tryptic-digest peptides were purified by reverse-phase h.p.l.c. and submitted to gas-phase sequencing. Up to eight sequences (79 amino acid residues) could be aligned with the human muscle enzyme sequence. Levels of identity range between 37 and 100%, indicating that, although human and yeast glycogen synthases probably share some conserved regions, significant differences in their primary structure should be expected.

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Processing of the precursor of protamine P2 in mouse. Peptide mapping and N-terminal sequence analysis of intermediates

Biochemical Journal ◽

10.1042/bj2770039 ◽

1991 ◽

Vol 277 (1) ◽

pp. 39-45 ◽

Cited By ~ 20

Author(s):

D Carré-Eusèbe ◽

F Lederer ◽

K H D Lê ◽

S M Elsevier

Keyword(s):

Amino Acid ◽

Sequence Analysis ◽

Peptide Mapping ◽

Chromatin Condensation ◽

Peptide Bond ◽

Chromosomal Protein ◽

Amino Acid Residues ◽

Terminal Sequence ◽

Vinylidene Difluoride ◽

N Terminus

Protamine P2, the major basic chromosomal protein of mouse spermatozoa, is synthesized as a precursor almost twice as long as the mature protein, its extra length arising from an N-terminal extension of 44 amino acid residues. This precursor is integrated into chromatin of spermatids, and the extension is processed during chromatin condensation in the haploid cells. We have studied processing in the mouse and have identified two intermediates generated by proteolytic cleavage of the precursor. H.p.l.c. separated protamine P2 from four other spermatid proteins, including the precursor and three proteins known to possess physiological characteristics expected of processing intermediates. Peptide mapping indicated that all of these proteins were structurally similar. Two major proteins were further purified by PAGE, transferred to poly(vinylidene difluoride) membranes and submitted to automated N-terminal sequence analysis. Both sequences were found within the deduced sequence of the precursor extension. The N-terminus of the larger intermediate, PP2C, was Gly-12, whereas the N-terminus of the smaller, PP2D, was His-21. Both processing sites involved a peptide bond in which the carbonyl function was contributed by an acidic amino acid.

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Mutational analysis of the channel and loop sequences of human ferritin H-chain

Biochemical Journal ◽

10.1042/bj2640381 ◽

1989 ◽

Vol 264 (2) ◽

pp. 381-388 ◽

Cited By ~ 69

Author(s):

S Levi ◽

A Luzzago ◽

F Franceschinelli ◽

P Santambrogio ◽

G Cesareni ◽

...

Keyword(s):

Amino Acid ◽

Iron Uptake ◽

Iron Oxidation ◽

Single Amino Acid ◽

Iron Core ◽

Amino Acid Residues ◽

Terminal Sequence ◽

Loop Sequence ◽

Human Ferritin ◽

Ferritin H

Human ferritin H-chain mutants were obtained by engineering the recombinant protein expressed by Escherichia coli. The mutagenesis were directed to the C-terminal sequence forming the hydrophobic channel, to the hydrophilic channel and to the loop sequence. The mutants were analysed for extent of expression, for stability, for capacity to incorporate iron and for kinetics of iron uptake and iron oxidation. Of the 22 mutants analysed only two with deletions of single residues in the loop sequence and one with deletion of the last 28 amino acid residues did not assemble into ferritin-like proteins. The other mutants assembled correctly and showed similar chemical/physical properties to the wild-type; they included duplication of an 18-amino acid-residue stretch, deletion of the last 22 and the last seven residues and various mutations of single amino acid residues. Two mutants with extensive alteration in the C-terminal sequence had a diminished thermostability associated with incapability to incorporate iron though they still catalysed iron oxidation. The mutants with alterations of the sequence around the hydrophilic channel showed diminished iron uptake and oxidation kinetics, together with a slightly larger apparent molecular size. The results indicate (i) that two of the sequences are important for ferritin assembly/stability, (ii) that the presence of the hydrophobic channel is essential for formation of the iron core and (iii) that the sites of iron interaction and the path of iron penetration into ferritin remain unidentified.

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Characterization of DNA-binding activity in the N-terminal domain of the DNA methyltransferase Dnmt3a

Biochemical Journal ◽

10.1042/bj20110241 ◽

2011 ◽

Vol 437 (1) ◽

pp. 141-148 ◽

Cited By ~ 24

Author(s):

Isao Suetake ◽

Yuichi Mishima ◽

Hironobu Kimura ◽

Young-Ho Lee ◽

Yuji Goto ◽

...

Keyword(s):

Dna Methylation ◽

Amino Acid ◽

Dna Binding ◽

Dna Methyltransferase ◽

Basic Amino Acid ◽

Binding Activity ◽

Amino Acid Residues ◽

Terminal Sequence ◽

Terminal Domain ◽

Dna Binding Activity

The Dnmt3a gene, which encodes de novo-type DNA methyltransferase, encodes two isoforms, full-length Dnmt3a and Dnmt3a2, which lacks the N-terminal 219 amino acid residues. We found that Dnmt3a showed higher DNA-binding and DNA-methylation activities than Dnmt3a2. The N-terminal sequence from residues 1 to 211 was able to bind to DNA, but could not distinguish methylated and unmethylated CpG. Its binding to DNA was inhibited by a major groove binder. Four basic amino acid residues, Lys51, Lys53, Arg177 and Arg179, in the N-terminal region were crucial for the DNA-binding activity. The ectopically expressed N-terminal sequence (residues 1–211) was localized in nuclei, whereas that harbouring mutations at the four basic amino acid residues was also detected in the cytoplasm. The DNA-methylation activity of Dnmt3a with the mutations was suppressed under physiological salt conditions, which is similar that of Dnmt3a2. In addition, ectopically expressed Dnmt3a with mutations, as well as Dnmt3a2, could not be retained efficiently in nuclei on salt extraction. We conclude that the DNA-binding activity of the N-terminal domain contributes to the DNA-methyltransferase activity via anchoring of the whole molecule to DNA under physiological salt conditions.

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