scholarly journals A structural and functional analogue of a Bowman–Birk-type protease inhibitor from Odorrana schmackeri

2017 ◽  
Vol 37 (2) ◽  
Author(s):  
Yuxin Wu ◽  
Qilin Long ◽  
Ying Xu ◽  
Shaodong Guo ◽  
Tianbao Chen ◽  
...  
Keyword(s):  
Protease Inhibitor ◽  
Frog Skin ◽  
Disulfide Bridge ◽  
Inhibitory Effect ◽  
Skin Secretion ◽  
Drug Discovery And Development ◽  
Trypsin Inhibitory Activity ◽  
Skin Secretions ◽  
Development Pipeline ◽  
First Time

Frog skin secretions contain complex peptidomes and peptidic protease inhibitors that are one of the biologically and structurally described groups of components. In the present study, by use of molecular ‘shotgun’ cloning and LC MS/MS fractionation sequencing, a novel Bowman–Birk-type heptadecapeptide (AALKGCWTKSIPPKPCF-amide), named Odorrana schmackeriTrypsin Inhibitor (OSTI), with a canonical Cys6–Cys16 disulfide bridge, was isolated and identified in piebald odorous frog (O. schmackeri) skin secretion. A synthetic replicate of OSTI-exhibited trypsin inhibitory activity with a Ki value of 0.3 ± 0.04 nM and also a tryptase inhibitory effect with a Ki of 2.5 ± 0.6 μM. This is the first time that this property has been reported for a peptide originating from amphibian sources. In addition, substituting lysine (K) with phenylalanine (F) at the presumed P1 position, completely abrogated the trypsin and tryptase inhibition, but produced a strong chymotrypsin inhibition with a Ki of 1.0 ± 0.1 μM. Thus, the specificity of this peptidic protease inhibitor could be optimized through modifying the amino acid residue at the presumed P1 position and this novel native OSTI, along with its analogue, [Phe9]-OSTI, have expanded the potential drug discovery and development pipeline directed towards alleviation of serine protease-mediated pathologies.

scholarly journals Discovery and Rational Design of a Novel Bowman-Birk Related Protease Inhibitor

Biomolecules ◽  
2019 ◽  
Vol 9 (7) ◽  
pp. 280 ◽  
Author(s):  
Yuxi Miao ◽  
Guanzhu Chen ◽  
Xinping Xi ◽  
Chengbang Ma ◽  
Lei Wang ◽  
...  
Keyword(s):  
Protease Inhibitor ◽  
Inhibitory Activity ◽  
Rational Design ◽  
Cdna Sequence ◽  
Cell Penetrating Peptide ◽  
Skin Secretion ◽  
Amphibian Skin ◽  
Skin Secretions ◽  
Inhibitor Drug ◽  
Potent Inhibitory Activity

Anuran amphibian skin secretions are a rich source of peptides, many of which represent novel protease inhibitors and can potentially act as a source for protease inhibitor drug discovery. In this study, a novel bioactive Bowman-Birk type inhibitory hexadecapeptide of the Ranacyclin family from the defensive skin secretion of the Fukien gold-striped pond frog, Pelophlax plancyi fukienesis, was successfully isolated and identified, named PPF-BBI. The primary structure of the biosynthetic precursor was deduced from a cDNA sequence cloned from a skin-derived cDNA library, which contains a consensus motif representative of the Bowman-Birk type inhibitor. The peptide was chemically synthesized and displayed a potent inhibitory activity against trypsin (Ki of 0.17 µM), as well as an inhibitory activity against tryptase (Ki of 30.73 µM). A number of analogues of this peptide were produced by rational design. An analogue, which substituted the lysine (K) at the predicted P1 position with phenylalanine (F), exhibited a potent chymotrypsin inhibitory activity (Ki of 0.851 µM). Alternatively, a more potent protease inhibitory activity, as well as antimicrobial activity, was observed when P16 was replaced by lysine, forming K16-PPF-BBI. The addition of the cell-penetrating peptide Tat with a trypsin inhibitory loop resulted in a peptide with a selective inhibitory activity toward trypsin, as well as a strong antifungal activity. This peptide also inhibited the growth of two lung cancer cells, H460 and H157, demonstrating that the targeted modifications of this peptide could effectively and efficiently alter its bioactivity.

scholarly journals Identification of a new myotropic decapeptide from the skin secretion of the red-eyed leaf frog, Agalychnis callidryas

PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0243326
Author(s):  
Yitian Gao ◽  
Renjie Li ◽  
Wenqing Yang ◽  
Mei Zhou ◽  
Lei Wang ◽  
...  
Keyword(s):  
Frog Skin ◽  
Binding Mode ◽  
Skin Secretion ◽  
Mature Peptide ◽  
Agalychnis Callidryas ◽  
C Terminus ◽  
Future Drug ◽  
Myotropic Activity ◽  
Skin Secretions ◽  
Drug Leads

Bradykinin-related peptides (BRPs) family is one of the most significant myotropic peptide families derived from frog skin secretions. Here, a novel BRP callitide was isolated and identified from the red-eyed leaf frog, Agalychnis callidryas, with atypical primary structure FRPAILVRPK-NH2. The mature peptide was cleaved N-terminally at a classic propeptide convertase cleavage site (-KR-) and at the C-terminus an unusual -GKGKGK sequence was removed using the first G residue as an amide donor for the C-terminally-located K residue. Thereafter, the synthetic replicates of callitide were assessed the myotropic activity and showed a significant contraction of balder, with the 0.63 nM EC50 value, more potent than most discovered myotropic peptides. The binding mode was further speculated by molecular docking and stimulation. The result indicated that the C-terminal of callitide might selectively bind to bradykinin receptor B2 (BKRB2). Further investigation of the callitide needs to be done in the future to be exploited as potential future drug leads.

scholarly journals Antibiofilm Activities in Skin Secretions of Malaysian Frogs

2019 ◽  
Vol 7 (4.14) ◽  
pp. 14
Author(s):  
A Abdul-Aziz ◽  
M S Razali ◽  
W M S Wan Azmi ◽  
Z A Rahman ◽  
M F F Abdullah
Keyword(s):  
Biofilm Formation ◽  
High Performance ◽  
Frog Skin ◽  
Maturation Stage ◽  
Antibiofilm Activity ◽  
Skin Secretion ◽  
Biofilm Inhibition ◽  
Maturation Stages ◽  
Skin Secretions ◽  
Extracellular Structures

Biofilms are extracellular structures formed by many species of bacteria that attached on various surfaces. The formation of biofilms is responsible for many health problems as they are very difficult to treat effectively due to enhanced resistant to antibiotics. Frog skin secretions contain bioactive compounds that may exhibit antibiofilms properties against biofilms infection. This study aims to screen the presence of antibiofilm activities from lyophilized skin secretions collected from Malaysian frogs. In this study, lyophilized skin secretions from four species of Malaysian frogs were tested for biofilm inhibition activities against the bacteria Staphylococcus epidermidis, ATCC 35984. Antibiofilm activity was tested at three different stages of biofilm formation which were biofilm maturation, biofilm attachment and biofilm dispersion. Biofilm treated with skin secretion from Fejervarya multistriata showed the presence of antibiofilm activities of 45 % which was able to reduce biofilm formation to 55 % at the attachment stage. Similarly, the biofilm formation at the maturation stage was reduced to 65.8 % only. No antibiofilm activities of Fejervarya multistriata was detected at the dispersion stage. The sample showing positive results was fractionated using column chromatography and further purified by C18 reverse phase high-performance liquid chromatography column (HPLC). Mass spectroscopy revealed the presence of a single peptide. In conclusion, frog skin secretion of F. multistriata contains peptide that inhibits biofilm at both attachment and maturation stages. 

PRELIMINARY STUDY OF ANTIMICROBIAL ACTIVITY OF THE SKIN SECRETIONS OF MALAYSIAN FROGS

2016 ◽  
Vol 78 (6-5) ◽  
Author(s):  
Zainon A. Rahman ◽  
Mohamad Faiz Foong Abdullah ◽  
Wan Mohamad Syahmin Wan Azmi ◽  
Aziyah Abdul-Aziz
Keyword(s):  
Antimicrobial Activity ◽  
Bioactive Compounds ◽  
Frog Skin ◽  
Antimicrobial Properties ◽  
Antimicrobial Effect ◽  
Skin Secretion ◽  
Gram Positive Bacteria ◽  
Microbial Infections ◽  
Skin Secretions ◽  
Preliminary Study

Bioactive compounds that exhibit antimicrobial properties in frogs are parts of the animal defense against microbial infections.  The lyophilized frog’s skin secretions containing varies bioactive compounds were subjected to screen for their antimicrobial activity. This study was conducted as part of an effort on the search of antimicrobial peptides (AMPs) profiles of Malaysian frogs. The results indicate the collected frog skin secretion has antimicrobial effect against Gram-negative and Gram-positive bacteria.  BLAST and standard phylogenetics were used to establish a preliminary identity of the frog samples.

scholarly journals Balteatide: A Novel Antimicrobial Decapeptide from the Skin Secretion of the Purple-Sided Leaf Frog,Phyllomedusa baltea

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Lilin Ge ◽  
Xiaole Chen ◽  
Chengbang Ma ◽  
Mei Zhou ◽  
Xinping Xi ◽  
...  
Keyword(s):  
Amino Acid ◽  
Rational Design ◽  
Frog Skin ◽  
Biologically Active ◽  
Single Amino Acid ◽  
Haemolytic Activity ◽  
Amino Acid Difference ◽  
Amino Acid Residues ◽  
Skin Secretion ◽  
Skin Secretions

The skin secretions of Neotropical phyllomedusine leaf frogs have proven to be a rich source of biologically active peptides, including antimicrobials. The major families of antimicrobial peptides (AMPs) reported are the dermaseptins and phylloseptins and the minor families are the dermatoxins, phylloxins, plasticins, distinctins, and medusins. Here, we report a novel AMP of 10 amino acid residues (LRPAILVRIKamide), named balteatide, from the skin secretion of wild Peruvian purple-sided leaf frogs,Phyllomedusa baltea. Balteatide was found to exhibit a 90% sequence identity with sauvatide, a potent myotropic peptide from the skin secretion ofPhyllomedusa sauvagei. However, despite both peptides exhibiting only a single amino acid difference (I/T at position 9), sauvatide is devoid of antimicrobial activity and balteatide is devoid of myotropic activity. Balteatide was found to have differential activity against the Gram-positive bacterium,Staphylococcus aureus; the Gram-negative bacterium,Escherichia coli; and the yeast,Candida albicans, and unusual for phyllomedusine frog skin AMPs, was most potent (MIC 32 mg/L) against the yeast. Balteatide was also devoid of haemolytic activity up to concentrations of 512 mg/L. Phyllomedusine frog skin secretions thus continue to provide novel AMPs, some of which may provide templates for the rational design of new classes of anti-infective therapeutics.

scholarly journals A Novel Kunitzin-Like Trypsin Inhibitor Isolated from Defensive Skin Secretion of Odorrana versabilis

Biomolecules ◽  
2019 ◽  
Vol 9 (7) ◽  
pp. 254 ◽  
Author(s):  
Yanjing Dong ◽  
Daning Shi ◽  
Yuan Ying ◽  
Xinping Xi ◽  
Xiaoling Chen ◽  
...  
Keyword(s):  
Trypsin Inhibitor ◽  
Inhibitory Activity ◽  
The Novel ◽  
Skin Secretion ◽  
Amphibian Skin ◽  
Sequence Alignments ◽  
Molecular Weights ◽  
Trypsin Inhibitory Activity ◽  
Skin Secretions ◽  
Potent Inhibitory Activity

Protease inhibitors that were identified from amphibian skin secretions with low molecular weights and potent inhibitory activity were thought to be potential candidates for novel peptide drugs. Here, a novel peptide with trypsin inhibitory activity was found in the skin secretion of the Chinese bamboo leaf odorous frog, Odorrana versabilis. Based on the sequence alignments of sequencing results, the novel peptide (ALKYPFRCKAAFC) was named as Kunitzin-OV. The synthetic replicate of Kunitzin-OV was subjected to a series of functional assays, and it exhibited a trypsin inhibitory activity with a Ki value of 3.042 µM, whereas, when Lys-9 at P1 position was substituted by Phe, trypsin inhibitory activity was undetected and the chymotrypsin inhibitory activity was optimized with a Ki value of 2.874 µM. However, its protease-binding loop was catabolized by trypsin during the trypsin cleavage test. In conclusion, Kunizin-OV is a novel peptide with trypsin inhibitory activity as a member of kunitzins, which is a non-typical Kunitz-like trypsin inhibitor with a highly conserved reactive site (K-A) and quite a short sequence.

scholarly journals Bufadienolides from the Skin Secretions of the Neotropical Toad Rhinella alata (Anura: Bufonidae): Antiprotozoal Activity against Trypanosoma cruzi

Molecules ◽  
2021 ◽  
Vol 26 (14) ◽  
pp. 4217
Author(s):  
Candelario Rodriguez ◽  
Roberto Ibáñez ◽  
Luis Mojica ◽  
Michelle Ng ◽  
Carmenza Spadafora ◽  
...  
Keyword(s):  
Vero Cells ◽  
Chemical Diversity ◽  
Hydroxyl Group ◽  
Antitrypanosomal Activity ◽  
The Family ◽  
Toad Venom ◽  
Skin Secretions ◽  
Pharmacological Potential ◽  
First Time ◽  
Mcf 7

Toads in the family Bufonidae contain bufadienolides in their venom, which are characterized by their chemical diversity and high pharmacological potential. American trypanosomiasis is a neglected disease that affects an estimated 8 million people in tropical and subtropical countries. In this research, we investigated the chemical composition and antitrypanosomal activity of toad venom from Rhinella alata collected in Panama. Structural determination using mass spectrometry (MS) and nuclear magnetic resonance (NMR) spectroscopy led to the identification of 10 bufadienolides. Compounds identified include the following: 16β-hydroxy-desacetyl-bufotalin-3-adipoyl-arginine ester (1), bufotalin (2), 16β-hydroxy-desacetyl-bufotalin-3-pimeloyl-arginine ester (3), bufotalin-3-pimeloyl-arginine ester (4), 16β-hydroxy-desacetyl-bufotalin-3-suberoyl-arginine ester (5), bufotalin-3-suberoyl-arginine ester (6), cinobufagin-3-adipoyl-arginine ester (7), cinobufagin-3-pimeloyl-arginine ester (8), cinobufagin-3-suberoyl-arginine ester (9), and cinobufagin (10). Among these, three new natural products, 1, 3, and 5, are described, and compounds 1–10 are reported for the first time in R. alata. The antitrypanosomal activity assessed in this study revealed that the presence of an arginyl-diacid attached to C-3, and a hydroxyl group at C-14 in the structure of bufadienolides that is important for their biological activity. Bufadienolides showed cytotoxic activity against epithelial kidney Vero cells; however, bufagins (2 and 10) displayed low mammalian cytotoxicity. Compounds 2 and 10 showed activity against the cancer cell lines MCF-7, NCI-H460, and SF-268.

scholarly journals Brassinolide Enhances the Level of Brassinosteroids, Protein, Pigments, and Monosaccharides in Wolffia arrhiza Treated with Brassinazole

Plants ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1311
Author(s):  
Magdalena Chmur ◽  
Andrzej Bajguz
Keyword(s):  
Plant Growth ◽  
Growth And Development ◽  
Inhibitory Effect ◽  
Dependent Manner ◽  
Plant Growth And Development ◽  
Concentration Dependent Manner ◽  
Spectrophotometric Methods ◽  
Synthetic Inhibitor ◽  
Wolffia Arrhiza ◽  
First Time

Brassinolide (BL) represents brassinosteroids (BRs)—a group of phytohormones that are essential for plant growth and development. Brassinazole (Brz) is as a synthetic inhibitor of BRs’ biosynthesis. In the present study, the responses of Wolffia arrhiza to the treatment with BL, Brz, and the combination of BL with Brz were analyzed. The analysis of BRs and Brz was performed using LC-MS/MS. The photosynthetic pigments (chlorophylls, carotenes, and xanthophylls) levels were determined using HPLC, but protein and monosaccharides level using spectrophotometric methods. The obtained results indicated that BL and Brz influence W. arrhiza cultures in a concentration-dependent manner. The most stimulatory effects on the growth, level of BRs (BL, 24-epibrassinolide, 28-homobrassinolide, 28-norbrassinolide, catasterone, castasterone, 24-epicastasterone, typhasterol, and 6-deoxytyphasterol), and the content of pigments, protein, and monosaccharides, were observed in plants treated with 0.1 µM BL. Whereas the application of 1 µM and 10 µM Brz caused a significant decrease in duckweed weight and level of targeted compounds. Application of BL caused the mitigation of the Brz inhibitory effect and enhanced the BR level in duckweed treated with Brz. The level of BRs was reported for the first time in duckweed treated with BL and/or Brz.

scholarly journals Unravelling the Skin Secretion Peptides of the Gliding Leaf Frog, Agalychnis spurrelli (Hylidae)

Biomolecules ◽  
2019 ◽  
Vol 9 (11) ◽  
pp. 667 ◽  
Author(s):  
Carolina Proaño-Bolaños ◽  
Ailín Blasco-Zúñiga ◽  
José Rafael Almeida ◽  
Lei Wang ◽  
Miguel Angel Llumiquinga ◽  
...  
Keyword(s):  
Inhibitory Concentration ◽  
Structural Diversity ◽  
Biologically Active ◽  
Biological Characterization ◽  
Skin Secretion ◽  
Peptide Profile ◽  
Active Chemical ◽  
Molecular Masses ◽  
Skin Secretions

Frog skin secretions contain medically-valuable molecules, which are useful for the discovery of new biopharmaceuticals. The peptide profile of the skin secretion of Agalychnis spurrelli has not been investigated; therefore, the structural and biological characterization of its compounds signify an inestimable opportunity to acquire new biologically-active chemical scaffolds. In this work, skin secretion from this amphibian was analysed by molecular cloning and tandem mass spectrometry. Although the extent of this work was not exhaustive, eleven skin secretion peptides belonging to five peptide families were identified. Among these, we report the occurrence of two phyllokinins, and one medusin-SP which were previously reported in other related species. In addition, eight novel peptides were identified, including four dermaseptins, DRS-SP2 to DRS-SP5, one phylloseptin-SP1, and three orphan peptides. Phylloseptin-SP1 and dermaseptins-SP2 were identified in HPLC fractions based on their molecular masses determined by MALDI-TOF MS. Among the antimicrobial peptides, dermaseptin-SP2 was the most potent, inhibiting Escherichia coli, Staphylococcus aureus, and ORSA with a minimum inhibitory concentration (MIC) of 2.68 μM, and Candida albicans with an MIC of 10.71 μM, without haemolytic effects. The peptides described in this study represent but a superficial glance at the considerable structural diversity of bioactive peptides produced in the skin secretion of A. spurrelli.

scholarly journals A novel role for WAVE1 in controlling actin network growth rate and architecture

2015 ◽  
Vol 26 (3) ◽  
pp. 495-505 ◽  
Author(s):  
Meredith O. Sweeney ◽  
Agnieszka Collins ◽  
Shae B. Padrick ◽  
Bruce L. Goode
Keyword(s):  
Actin Filament ◽  
Specific Activity ◽  
Inhibitory Effect ◽  
Actin Network ◽  
Inhibitory Effects ◽  
Cellular Functions ◽  
Network Growth ◽  
Assembly Kinetics ◽  
Filament Networks ◽  
First Time

Branched actin filament networks in cells are assembled through the combined activities of Arp2/3 complex and different WASP/WAVE proteins. Here we used TIRF and electron microscopy to directly compare for the first time the assembly kinetics and architectures of actin filament networks produced by Arp2/3 complex and dimerized VCA regions of WAVE1, WAVE2, or N-WASP. WAVE1 produced strikingly different networks from WAVE2 or N-WASP, which comprised unexpectedly short filaments. Further analysis showed that the WAVE1-specific activity stemmed from an inhibitory effect on filament elongation both in the presence and absence of Arp2/3 complex, which was observed even at low stoichiometries of WAVE1 to actin monomers, precluding an effect from monomer sequestration. Using a series of VCA chimeras, we mapped the elongation inhibitory effects of WAVE1 to its WH2 (“V”) domain. Further, mutating a single conserved lysine residue potently disrupted WAVE1's inhibitory effects. Taken together, our results show that WAVE1 has unique activities independent of Arp2/3 complex that can govern both the growth rates and architectures of actin filament networks. Such activities may underlie previously observed differences between the cellular functions of WAVE1 and WAVE2.

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