Protein chemotaxonomy of genus Datura: Identical amino acid sequence of ferredoxin from two varieties of Datura stramonium

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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Isolation and characterization of a cDNA clone coding for the calcium-binding subunit of calcineurin from bovine brain: An identical amino acid sequence to the human protein

DNA Sequence ◽

10.3109/10425179409020857 ◽

1994 ◽

Vol 4 (5) ◽

pp. 313-318 ◽

Cited By ~ 3

Author(s):

Cheryl E. Nargang ◽

Drell A. Bottorff ◽

Kazuo Adachi

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Cdna Clone ◽

Calcium Binding ◽

Bovine Brain ◽

Human Protein ◽

Isolation And Characterization ◽

Identical Amino Acid Sequence ◽

Binding Subunit

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Characteristics of Classical Swine Fever Virus Variants Derived from Live Attenuated GPE− Vaccine Seed

Viruses ◽

10.3390/v13081672 ◽

2021 ◽

Vol 13 (8) ◽

pp. 1672

Author(s):

Taksoo Kim ◽

Loc Tan Huynh ◽

Shizuka Hirose ◽

Manabu Igarashi ◽

Takahiro Hiono ◽

...

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Type I Interferon ◽

Classical Swine Fever ◽

Amino Acid Substitutions ◽

Type I ◽

Live Attenuated Vaccine ◽

Marker Vaccine ◽

Seed Availability ◽

Identical Amino Acid Sequence

The GPE− strain is a live attenuated vaccine for classical swine fever (CSF) developed in Japan. In the context of increasing attention for the differentiating infected from vaccinated animals (DIVA) concept, the achievement of CSF eradication with the GPE− proposes it as a preferable backbone for a recombinant CSF marker vaccine. While its infectious cDNA clone, vGPE−, is well characterized, 10 amino acid substitutions were recognized in the genome, compared to the original GPE− vaccine seed. To clarify the GPE− seed availability, this study aimed to generate and characterize a clone possessing the identical amino acid sequence to the GPE− seed. The attempt resulted in the loss of the infectious GPE− seed clone production due to the impaired replication by an amino acid substitution in the viral polymerase NS5B. Accordingly, replication-competent GPE− seed variant clones were produced. Although they were mostly restricted to propagate in the tonsils of pigs, similarly to vGPE−, their type I interferon-inducing capacity was significantly lower than that of vGPE−. Taken together, vGPE− mainly retains ideal properties for the CSF vaccine, compared with the seed variants, and is probably useful in the development of a CSF marker vaccine.

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Urocortin is the principal ligand for the corticotrophin-releasing factor binding protein in the ovine brain with no evidence for a sauvagine-like peptide

Journal of Molecular Endocrinology ◽

10.1677/jme.0.0240053 ◽

2000 ◽

Vol 24 (1) ◽

pp. 53-63 ◽

Cited By ~ 13

Author(s):

SM Baigent ◽

PJ Lowry

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Binding Protein ◽

Peptide Ligands ◽

Terminal Amino Acid ◽

Factor Binding ◽

Corticotrophin Releasing Factor ◽

Identical Amino Acid Sequence ◽

Successful Technique ◽

Terminal Amino

To purify novel ligands for the corticotrophin-releasing factor binding protein (CRF-BP) from ovine brain, whole brain was homogenised in methanol and the supernatant extracted on Sep-pak C18 cartridges followed by a preliminary HPLC step. Three peaks of ovine CRF-BP ligand activity were detected in the HPLC fractions, the first two of which were also detected by a specific corticotrophin-releasing factor two-site immunoradiometric assay, the third peak being detected by a human CRF-BP ligand assay, which will not detect ovine CRF. Human CRF-BP ligand-containing fractions were further purified by affinity chromatography on a human recombinant CRF-BP column with two additional HPLC steps. The human CRF-BP ligand was found to: (a) possess a molecular mass of 4707 Daltons, (b) have an N-terminal amino acid sequence (5 residues) identical to rat urocortin, (c) be detected by a specific urocortin radioimmunoassay, (d) have high affinity for both the human and ovine CRF-BPs and (e) be present in many regions of the ovine brain. Additionally, a 300 bp cDNA fragment sharing 83% homology with the rat urocortin gene was cloned from ovine brain, the product of which was predicted to have an identical amino acid sequence to that of rat urocortin. These pieces of information confirmed the identity of the human CRF-BP ligand as an ovine urocortin. The specially developed CRF-BP ligand assays showed that the rank orders of affinity of the CRF family members for human CRF-BP were: carp urotensin-1>>human CRF=rat/ovine urocortin>human urocortin>>frog sauvagine>>ovine CRF, and those for the ovine CRF-BP were: carp urotensin-1> human CRF=rat/ovine urocortin>human urocortin> frog sauvagine>>ovine CRF. This study describes a successful technique for the purification and detection of peptide ligands for the CRF-BP. We conclude that urocortin is the principal ligand for the CRF-BP in ovine brain and we could find no evidence for a centrally located mammalian sauvagine-like peptide.

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