Hernández-Arvizu Edwin Esaú, Silis-Moreno Teresa Monserrat, García-Arredondo José Alejandro, Rodríguez-Torres Angelina, Cervantes-Chávez José Antonio, Mosqueda Juan
Immunology and Vaccine Research Laboratory, Natural Sciences College, Autonomous University of Queretaro, Queretaro 76230, Mexico.
Ph.D. Program in Natural Sciences, Natural Sciences College, Autonomous University of Queretaro, Queretaro 76230, Mexico.
Microorganisms. 2023 Nov 15;11(11):2778. doi: 10.3390/microorganisms11112778.
Anti-microbial peptides play a vital role in the defense mechanisms of various organisms performing functions that range from the elimination of microorganisms, through diverse mechanisms, to the modulation of the immune response, providing protection to the host. Among these peptides, cathelicidins, a well-studied family of anti-microbial peptides, are found in various animal species, including reptiles. Due to the rise in anti-microbial resistance, these compounds have been suggested as potential candidates for developing new drugs. In this study, we identified and characterized a cathelicidin-like peptide called Aquiluscidin (Aq-CATH) from transcripts obtained from the skin and oral mucosa of the Querétaro's dark rattlesnake, . The cDNA was cloned, sequenced, and yielded a 566-base-pair sequence. Using bioinformatics, we predicted that the peptide precursor contains a signal peptide, a 101-amino-acid conserved cathelin domain, an anionic region, and a 34-amino-acid mature peptide in the C-terminal region. Aq-CATH and a derived 23-amino-acid peptide (Vcn-23) were synthesized, and their anti-microbial activity was evaluated against various species of bacteria in in vitro assays. The minimal inhibitory concentrations against bacteria ranged from 2 to 8 μg/mL for both peptides. Furthermore, at concentrations of up to 50 μM, they exhibited no significant hemolytic activity (<2.3% and <1.2% for Aquiluscidin and Vcn-23, respectively) against rat erythrocytes and displayed no significant cytotoxic activity at low concentrations (>65% cell viability at 25 µM). Finally, this study represents the first identification of an antimicrobial peptide in , which belongs to the cathelicidin family and exhibits the characteristic features of these peptides. Both Aq-CATH and its derived molecule, Vcn-23, displayed remarkable inhibitory activity against all tested bacteria, highlighting their potential as promising candidates for further antimicrobial research.
抗菌肽在各种生物体的防御机制中发挥着至关重要的作用,其功能从通过多种机制消除微生物到调节免疫反应,为宿主提供保护。在这些肽中,cathelicidins是一类经过充分研究的抗菌肽家族,存在于包括爬行动物在内的各种动物物种中。由于抗菌耐药性的增加,这些化合物被认为是开发新药的潜在候选物。在本研究中,我们从克雷塔罗黑腹响尾蛇的皮肤和口腔黏膜获得的转录本中鉴定并表征了一种名为Aquiluscidin(Aq-CATH)的类cathelicidin肽。对该cDNA进行了克隆、测序,得到了一个566个碱基对的序列。利用生物信息学,我们预测该肽前体包含一个信号肽、一个101个氨基酸的保守cathelin结构域、一个阴离子区域以及C末端区域的一个34个氨基酸的成熟肽。合成了Aq-CATH和一种衍生的23个氨基酸的肽(Vcn-23),并在体外试验中评估了它们对各种细菌的抗菌活性。两种肽对细菌的最低抑菌浓度范围为2至8μg/mL。此外,在浓度高达50μM时,它们对大鼠红细胞均未表现出明显的溶血活性(Aquiluscidin和Vcn-23分别<2.3%和<1.2%),并且在低浓度下(25μM时细胞活力>65%)未表现出明显的细胞毒性活性。最后,本研究首次在克雷塔罗黑腹响尾蛇中鉴定出一种抗菌肽,它属于cathelicidin家族并具有这些肽的特征。Aq-CATH及其衍生分子Vcn-23对所有测试细菌均表现出显著的抑制活性,突出了它们作为进一步抗菌研究的有前途候选物的潜力。