Identification of cathelicidin gene from Hoplobatrachus rugulosus and the antioxidant capacity of PC29 peptide

Abstract Cathelicidins, a group of vertebrate multifunctional molecules, play a role in innate immunity. In this study, a cathelicidin was identified from the lungs of frogs, Hoplobatrachus rugulosus. A 474 base pairs (bp) complementary DNA (cDNA) sequence encoded a 157 amino acid residue prepropeptide of H. rugulosus cathelicidin (cathelicidin-HR), which consisting of a 20-residue signal peptide sequence, a 108-residue cathelin region, and a 29-residue cathelicidin-HR peptide. Amino acid sequence alignment and cladogram analysis illustrated that cathelicidin-HR have a high degree of similarity to further amphibian cathelicidins. The cathelicidin-HR peptide displays very low antimicrobial activity but exhibits dose-dependent antioxidant activity. Moreover, this peptide expresses DNA damage inhibition against UV/H2O2-induction. The molecular docking indicated that DNA damage protection of cathelicidin-HR might occur via DNA-peptide complex formation. This is the first amphibian cathelicidin peptide that possesses DNA damage inhibitory activity which might play a crucial role in oxidative stress.

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Identification of cathelicidin gene from Hoplobatrachus rugulosus and the antioxidant capacity of PC29 peptide

10.21203/rs.3.rs-521559/v1 ◽

2021 ◽

Author(s):

Anupong Tankrathok ◽

Chutima Karnmongkol ◽

Arpaporn Punpad ◽

Piyachat Wiriyaumpaiwong ◽

Nattapong Srisam ◽

...

Keyword(s):

Amino Acid ◽

Peptide Sequence ◽

Base Pairs ◽

Amphibian Species ◽

Signal Peptide Sequence ◽

Microbial Invasion ◽

Multifunctional Molecules ◽

Functional Peptide ◽

High Degree ◽

Dose Dependent

Abstract Cathelicidins, a group of vertebrate multifunctional molecules, play a role in innate immunity. Cathelicidins are antimicrobial peptides (AMPs) that are involved in protection against microbial invasion. Presently, cathelicidin peptides have been identified from only 14 amphibian species. In the study, a novel cathelicidin was identified from the lungs of frogs, Hoplobatrachus rugulosus. A 474 base pairs (bp) complementary DNA (cDNA) sequence encoded a 157 amino acid residue prepropeptide of H. rugulosus cathelicidin (cathelicidin-HR), which consisting of a 20-residue signal peptide sequence, a 108-residue cathelin region, and a 29-residue cathelicidin peptide (PC29). Amino acid sequence alignment and cladogram analysis illustrated that cathelicidin-HR have a high degree of similarity to further amphibian cathelicidins. The PC29 peptide displays antimicrobial activity only against Bacillus subtilis and Enterococcus faecalis. However, the PC29 peptide performed dose-dependent antioxidant activity. This is the first cathelicidin antioxidant peptide identified from the lung which provided a template for the development of potent bi-functional peptide therapeutic agents.

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Amino acid sequence of the penicillin-binding protein/dd-peptidase of Streptomyces K15. Predicted secondary structures of the low Mr penicillin-binding proteins of class A

Biochemical Journal ◽

10.1042/bj2790223 ◽

1991 ◽

Vol 279 (1) ◽

pp. 223-230 ◽

Cited By ~ 13

Author(s):

P Palomeque-Messia ◽

S Englebert ◽

M Leyh-Bouille ◽

M Nguyen-Distèche ◽

C Duez ◽

...

Keyword(s):

Amino Acid ◽

Amino Acid Residue ◽

Active Sites ◽

Binding Protein ◽

Secondary Structures ◽

Peptide Sequence ◽

Penicillin Binding Protein ◽

Signal Peptide Sequence ◽

Beta Lactamases ◽

Class A

The low-Mr penicillin-binding protein (PBP)/DD-transpeptidase of Streptomyces K15 is synthesized in the form of a 291-amino acid-residue precursor possessing a cleavable 29-amino acid-residue signal peptide. Sequence-similarity searches and hydrophobic-cluster analysis show that the Streptomyces K15 enzyme, the Escherichia coli PBPs/DD-carboxy-peptidases 5 and 6, the Bacillus subtilis PBP/DD-carboxypeptidase 5 and the spoIIA product (a putative PBP involved in the sporulation of B. subtilis) are structurally related and form a distinct class A of low-Mr PBPs/DD-peptidases. The distribution of the hydrophobic clusters along the amino acid sequences also shows that the Streptomyces K15 PBP, and by extension the other PBPs of class A, have similarity in the polypeptide folding, with the beta-lactamases of class A, with as reference the Streptomyces albus G and Staphylococcus aureus beta-lactamases of known three-dimensional structure. This comparison allows one to predict most of the secondary structures in the PBPs and the amino acid motifs that define the enzyme active sites.

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Polysaccharide Lyase: Molecular Cloning, Sequencing, and Overexpression of the Xanthan Lyase Gene of Bacillus sp. Strain GL1

Applied and Environmental Microbiology ◽

10.1128/aem.67.2.713-720.2001 ◽

2001 ◽

Vol 67 (2) ◽

pp. 713-720 ◽

Cited By ~ 14

Author(s):

Wataru Hashimoto ◽

Hikaru Miki ◽

Noriaki Tsuchiya ◽

Hirokazu Nankai ◽

Kousaku Murata

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Molecular Mass ◽

Signal Peptide ◽

Streptomyces Griseus ◽

Peptide Sequence ◽

Terminal Amino Acid ◽

Signal Peptide Sequence ◽

Terminal Amino ◽

Terminal Amino Acid Sequence

ABSTRACT When grown on xanthan as a carbon source, the bacteriumBacillus sp. strain GL1 produces extracellular xanthan lyase (75 kDa), catalyzing the first step of xanthan depolymerization (H. Nankai, W. Hashimoto, H. Miki, S. Kawai, and K. Murata, Appl. Environ. Microbiol. 65:2520–2526, 1999). A gene for the lyase was cloned, and its nucleotide sequence was determined. The gene contained an open reading frame consisting of 2,793 bp coding for a polypeptide with a molecular weight of 99,308. The polypeptide had a signal peptide (2 kDa) consisting of 25 amino acid residues preceding the N-terminal amino acid sequence of the enzyme and exhibited significant homology with hyaluronidase of Streptomyces griseus(identity score, 37.7%). Escherichia coli transformed with the gene without the signal peptide sequence showed a xanthan lyase activity and produced intracellularly a large amount of the enzyme (400 mg/liter of culture) with a molecular mass of 97 kDa. During storage at 4°C, the purified enzyme (97 kDa) from E. coli was converted to a low-molecular-mass (75-kDa) enzyme with properties closely similar to those of the enzyme (75 kDa) fromBacillus sp. strain GL1, specifically in optimum pH and temperature for activity, substrate specificity, and mode of action. Logarithmically growing cells of Bacillus sp. strain GL1 on the medium with xanthan were also found to secrete not only xanthan lyase (75 kDa) but also a 97-kDa protein with the same N-terminal amino acid sequence as that of xanthan lyase (75 kDa). These results suggest that, in Bacillus sp. strain GL1, xanthan lyase is first synthesized as a preproform (99 kDa), secreted as a precursor (97 kDa) by a signal peptide-dependent mechanism, and then processed into a mature form (75 kDa) through excision of a C-terminal protein fragment with a molecular mass of 22 kDa.

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Characterisation and functional analysis of canine TLR5

Innate Immunity ◽

10.1177/1753425920901862 ◽

2020 ◽

Vol 26 (6) ◽

pp. 451-458

Author(s):

Aihua Zhu ◽

Lingling Wei ◽

Sujuan Hu ◽

Cheng Yang ◽

Caifa Chen ◽

...

Keyword(s):

Amino Acid ◽

Amino Acid Sequence ◽

Salmonella Enteritidis ◽

Transmembrane Domain ◽

Immunofluorescence Assay ◽

Peptide Sequence ◽

Signal Peptide Sequence ◽

Reading Frame ◽

Serovar Typhimurium ◽

Receptor Domain

In this study, we characterised the single exon TLR5 gene of the Chinese rural dog. Sequence analysis revealed a 2577 nucleotide-long open reading frame of canine TLR5, encoding an 858 amino acid-long protein. The putative amino acid sequence of canine TLR5 consisted of a signal peptide sequence, 15 LRR domains, a LRR C-terminal domain, a transmembrane domain and an intracellular Toll-IL-1 receptor domain. The amino acid sequence of the canine TLR5 protein shared 95.4% identity with vulpine, 72.2% with feline and 64.7% with human TLR5. Plasmids expressing canine TLR5 and NF-κB-luciferase were constructed and transfected into HEK293T cells. Expression was confirmed by indirect immunofluorescence assay. These HEK293T cells transfected with the canine TLR5- and NF-κB-luciferase plasmids signiﬁcantly responded to ﬂagellin from Salmonella enteritidis serovar Typhimurium, indicating that it is a functional TLR5 homolog. In response to stimulation with Salmonella enteritidis, the level of TLR5 mRNA significantly increased over the control in PBMCs at 4 h. The levels of IL-8, IL-6 and IL-1β also increased after exposure. The highest levels of TLR5, IL-8 and IL-1β expression were detected at 8, 4 and 12 h after stimulation, respectively. These results imply that the expression of canine TLR5 may participate in the immune response against bacterial pathogens.

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Molecular Cloning of Bovine Amelogenin cDna

Advances in Dental Research ◽

10.1177/08959374870010022001 ◽

1987 ◽

Vol 1 (2) ◽

pp. 293-297 ◽

Cited By ~ 47

Author(s):

H. Shimokawa ◽

Y. Ogata ◽

S. Sasaki ◽

M.E. Sobel ◽

C.I. Mcquillan ◽

...

Keyword(s):

Amino Acid ◽

Molecular Cloning ◽

Peptide Sequence ◽

Cdna Expression Library ◽

Cdna Clones ◽

Coding Region ◽

Expression Library ◽

Amino Acid Homology ◽

Signal Peptide Sequence ◽

Cloned Cdna

Molecular cloning of a bovine amelogenin cDNA was accomplished by construction of a cDNA expression library (λgt11 cDNA library) from the bovine ameloblast mRNA and then screening of the library with antibodies to bovine amelogenins. The complete primary structure of an amelogenin was deduced from cloned cDNA. One of the cDNA clones isolated from a bovine ameloblast phage λgt11 library had an 864-base-pair-long insert that encoded a protein with 216 amino acid residues. This cDNA clone appears to represent the complete coding region of amelogenin mRNA, including a putative AUG initiation codon and a signal peptide sequence. The predicted bovine amelogenin sequence has 87% amino acid homology with murine amelogenin.

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Amino acid sequence controls the self-assembled superstructure morphology of N-acetylated tri-β3-peptides

Pure and Applied Chemistry ◽

10.1515/pac-2015-0108 ◽

2015 ◽

Vol 87 (9-10) ◽

pp. 1021-1028 ◽

Cited By ~ 15

Author(s):

Rania S. Seoudi ◽

Annette Dowd ◽

Mark Del Borgo ◽

Ketav Kulkarni ◽

Patrick Perlmutter ◽

...

Keyword(s):

Amino Acid ◽

Self Assembly ◽

Far Infrared ◽

High Symmetry ◽

Amino Acid Residues ◽

Cross Sectional ◽

Self Assembling ◽

Afm Imaging ◽

High Degree ◽

Degree Of Similarity

AbstractPeptides based on unnatural β3-amino acids offer a versatile platform for the design of self-assembling nanostructures due to the folding stability of the 14-helix and the high symmetry of the side chains inherent in this geometry. We have previously described that N-terminal acetylation (Ac-) forms a supramolecular self-assembly motif that allows β3-peptides to assemble head-to-tail into a helical nanorod which then further bundles into hierarchical superstructures. Here we investigate the effect of the topography of the 14-helical nanorod on lateral self-assembly. Specifically, we report on the variations in the superstructure of three isomeric peptides comprising the same three β3-amino acid residues: β3-leucine (L), β3-isoleucine (I) β3-alanine (A) to give peptides Ac-β3[LIA], Ac-β3[IAL] and Ac-β3[ALI]. AFM imaging shows markedly different superstructures for the three peptides. Well defined synchrotron far-infrared spectra reveal uniform geometries with a high degree of similarity between the isomeric peptides in the amide modes of the 400–650 wavenumber range. Far-IR also confirms that the C-terminal carboxyl group is free in the assemblies, thus it is solvated in the dispersant. Hence, the differences in the superstructures formed by the fibers are defined primarily by van der Waals energy minimization between the varied cross sectional morphologies of the core nanorods.

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Identification and characterization of Saccharomyces cerevisiae yapsin 3, a new member of the yapsin family of aspartic proteases encoded by the YPS3 gene

Biochemical Journal ◽

10.1042/bj3390407 ◽

1999 ◽

Vol 339 (2) ◽

pp. 407-411 ◽

Cited By ~ 15

Author(s):

Vicki OLSEN ◽

Niamh X. CAWLEY ◽

Jakob BRANDT ◽

Michi EGEL-MITANI ◽

Y. Peng LOH

Keyword(s):

Saccharomyces Cerevisiae ◽

Amino Acid ◽

Cleavage Sites ◽

Basic Residue ◽

Aspartic Proteases ◽

N Terminus ◽

High Degree ◽

Identification And Characterization ◽

Degree Of Similarity

A new aspartic protease from Saccharomyces cerevisiae, with a high degree of similarity with yapsin 1 and yapsin 2 and a specificity for basic residue cleavage sites of prohormones, has been cloned. This enzyme was named yapsin 3. Expression of a C-terminally truncated non-membrane anchored yapsin 3 in yeast yielded a heterogeneous protein between 135–200 kDa which, upon treatment with endoglycosidase H, migrated as a 60 kDa form. Amino-acid analysis of the N-terminus of expressed yapsin 3 revealed two different N-terminal residues, serine-48 and phenylalanine-54, which followed a dibasic and a monobasic residue respectively. Cleavage of several prohormones by non-anchored yapsin 3 revealed a specificity distinct from that of yapsin 1.

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Structure of the human MLH1 N-terminus: implications for predisposition to Lynch syndrome

Acta Crystallographica Section F Structural Biology Communications ◽

10.1107/s2053230x15010183 ◽

2015 ◽

Vol 71 (8) ◽

pp. 981-985 ◽

Cited By ~ 11

Author(s):

Hong Wu ◽

Hong Zeng ◽

Robert Lam ◽

Wolfram Tempel ◽

Iain D. Kerr ◽

...

Keyword(s):

Genetic Testing ◽

Lynch Syndrome ◽

Mismatch Repair ◽

Base Pairs ◽

X Ray ◽

X Ray Crystallography ◽

N Terminus ◽

High Degree ◽

Degree Of Similarity

Mismatch repair prevents the accumulation of erroneous insertions/deletions and non-Watson–Crick base pairs in the genome. Pathogenic mutations in theMLH1gene are associated with a predisposition to Lynch and Turcot's syndromes. Although genetic testing for these mutations is available, robust classification of variants requires strong clinical and functional support. Here, the first structure of the N-terminus of human MLH1, determined by X-ray crystallography, is described. The structure shares a high degree of similarity with previously determined prokaryoticMLH1homologs; however, this structure affords a more accurate platform for the classification ofMLH1variants.

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Evaluation of Antioxidant and DNA Damage Protection Activity of the Hydroalcoholic Extract ofDesmostachya bipinnataL. Stapf

The Scientific World JOURNAL ◽

10.1155/2014/215084 ◽

2014 ◽

Vol 2014 ◽

pp. 1-8 ◽

Cited By ~ 17

Author(s):

Upendarrao Golla ◽

Solomon Sunder Raj Bhimathati

Keyword(s):

Dna Damage ◽

Antioxidant Activity ◽

Yeast Cells ◽

Dependent Manner ◽

Hydroalcoholic Extract ◽

Dose Dependent ◽

Protection Activity ◽

Dna Damage Protection ◽

Damage Protection

Desmostachya bipinnataStapf (Poaceae/Gramineae) is an official drug of ayurvedic pharmacopoeia. Various parts of this plant were used extensively in traditional and folklore medicine to cure various human ailments. The present study was aimed to evaluate the antioxidant and DNA damage protection activity of hydroalcoholic extract ofDesmostachya bipinnatabothin vitroandin vivo, to provide scientific basis for traditional usage of this plant. The extract showed significant antioxidant activity in a dose-dependent manner with an IC50value of264.18±3.47 μg/mL in H2O2scavenging assay and prevented the oxidative damage to DNA in presence of DNA damaging agent (Fenton’s reagent) at a concentration of 50 μg/mL. Also, the presence of extract protected yeast cells in a dose-dependent manner against DNA damaging agent (Hydroxyurea) in spot assay. Moreover, the presence of extract exhibited significant antioxidant activityin vivoby protecting yeast cells against oxidative stressing agent (H2O2). Altogether, the results of current study revealed thatDesmostachya bipinnatais a potential source of antioxidants and lends pharmacological credence to the ethnomedical use of this plant in traditional system of medicine, justifying its therapeutic application for free-radical-induced diseases.

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Study of the rat liver S-adenosylmethionine synthetase active site with 8-azido ATP

Biochemical Journal ◽

10.1042/bj3080565 ◽

1995 ◽

Vol 308 (2) ◽

pp. 565-571 ◽

Cited By ~ 7

Author(s):

H P Deigner ◽

J M Mato ◽

M A Pajares

Keyword(s):

Amino Acid ◽

Rat Liver ◽

Active Site ◽

Amino Acid Analysis ◽

Peptide Sequencing ◽

Case Identification ◽

Enzyme Sequence ◽

Adenosylmethionine Synthetase ◽

High Degree ◽

Degree Of Similarity

The active site of rat liver S-adenosylmethionine synthetase was studied using 8-azido ATP, a photolabile analogue of ATP. Both forms of the enzyme, tetramer and dimer, could be labelled by using concentrations of the analogue similar to the KmATP values for each form, 350 microM and 1 mM respectively. Labelling of both S-adenosylmethionine synthetase forms with 8-azido [alpha-32P]ATP, followed by tryptic digestion and purification by HPLC, afforded one specifically labelled peptide in each case. Identification of the labelled peptide by amino acid analysis and peptide sequencing, and comparison with the enzyme sequence, indicated that the same peptide (267-286) was modified in both enzyme forms. The results are discussed on the basis of the high degree of similarity that this peptide shows in all the known S-adenosylmethionine synthetase sequences.

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