Arpitha Ashok, Sebastin Santhosh M, Rohit A C, Girish K S, Vinod D, Aparna H S
DOS in Biotechnology, University of Mysore, Manasagangotri, Mysore, 570 006, India.
Department of Medical Biochemistry and Biophysics, Karolinska Institute, SE-171 77, Stockholm, Sweden.
Appl Biochem Biotechnol. 2017 Aug;182(4):1415-1432. doi: 10.1007/s12010-017-2407-6. Epub 2017 Feb 2.
Bioactive peptide research has experienced considerable therapeutic interest owing to varied physiological functions, efficacy in excretion, and tolerability of peptides. Colostrum is a rich natural source of bioactive peptides with many properties elucidated such as anti-thrombotic, anti-hypertensive, opioid, immunomodulatory, etc. In this study, a variant peptide derived from β-lactoglobulin from buffalo colostrum was evaluated for the anti-ophidian property by targeting snake venom metalloproteinases. These are responsible for rapid local tissue damages that develop after snakebite such as edema, hemorrhage, myonecrosis, and extracellular matrix degradation. The peptide identified by LC-MS/MS effectively neutralized hemorrhagic activity of the Echis carinatus venom in a dose-dependent manner. Histological examinations revealed that the peptide mitigated basement membrane degradation and accumulation of inflammatory leucocytes at the venom-injected site. Inhibition of proteolytic activity was evidenced in both casein and gelatin zymograms. Also, inhibition of fibrinolytic and fibrinogenolytic activities was seen. The UV-visible spectral study implicated Zn chelation, which was further confirmed by molecular docking and dynamic studies by assessing molecular interactions, thus implicating the probable mechanism for inhibition of venom-induced proteolytic and hemorrhagic activities. The present investigation establishes newer vista for the BLG-col peptide with anti-ophidian efficacy as a promising candidate for therapeutic interventions.
由于生物活性肽具有多种生理功能、排泄功效以及耐受性，其研究在治疗方面引起了广泛关注。初乳是生物活性肽丰富的天然来源，已阐明其具有许多特性，如抗血栓、抗高血压、类鸦片活性、免疫调节等。在本研究中，通过靶向蛇毒金属蛋白酶，评估了一种源自水牛初乳中β-乳球蛋白的变体肽的抗蛇毒特性。蛇毒金属蛋白酶会导致蛇咬后迅速出现局部组织损伤，如水肿、出血、肌坏死和细胞外基质降解。通过LC-MS/MS鉴定的该肽以剂量依赖方式有效中和了锯鳞蝰毒液的出血活性。组织学检查显示，该肽减轻了毒液注射部位的基底膜降解和炎症白细胞的聚集。酪蛋白和明胶酶谱均证明了蛋白水解活性受到抑制。此外，还观察到纤维蛋白溶解和纤维蛋白原溶解活性受到抑制。紫外可见光谱研究表明存在锌螯合作用，通过分子对接和动力学研究评估分子相互作用进一步证实了这一点，从而揭示了抑制毒液诱导的蛋白水解和出血活性的可能机制。本研究为具有抗蛇毒功效的BLG-col肽开辟了新的前景，使其成为治疗干预的有希望的候选物。
