Saviola Anthony J, Pla Davinia, Sanz Libia, Castoe Todd A, Calvete Juan J, Mackessy Stephen P
School of Biological Sciences, University of Northern Colorado, 501 20th Street, CB 92, Greeley, CO, 80639, USA; Instituto de Biomedicina de Valencia, C.S.I.C., Jaime Roig 11, 46010 Valencia, Spain.
Instituto de Biomedicina de Valencia, C.S.I.C., Jaime Roig 11, 46010 Valencia, Spain.
J Proteomics. 2015 May 21;121:28-43. doi: 10.1016/j.jprot.2015.03.015. Epub 2015 Mar 25.
Here we describe and compare the venomic and antivenomic characteristics of both neonate and adult Prairie Rattlesnake (Crotalus viridis viridis) venoms. Although both neonate and adult venoms contain unique components, similarities among protein family content were seen. Both neonate and adult venoms consisted of myotoxin, bradykinin-potentiating peptide (BPP), phospholipase A2 (PLA2), Zn(2+)-dependent metalloproteinase (SVMP), serine proteinase, L-amino acid oxidase (LAAO), cysteine-rich secretory protein (CRISP) and disintegrin families. Quantitative differences, however, were observed, with venoms of adults containing significantly higher concentrations of the non-enzymatic toxic compounds and venoms of neonates containing higher concentrations of pre-digestive enzymatic proteins such as SVMPs. To assess the relevance of this venom variation in the context of snakebite and snakebite treatment, we tested the efficacy of the common antivenom CroFab® for recognition of both adult and neonate venoms in vitro. This comparison revealed that many of the major protein families (SVMPs, CRISP, PLA2, serine proteases, and LAAO) in both neonate and adult venoms were immunodepleted by the antivenom, whereas myotoxins, one of the major toxic components of C. v. viridis venom, in addition to many of the small peptides, were not efficiently depleted by CroFab®. These results therefore provide a comprehensive catalog of the venom compounds present in C. v. viridis venom and new molecular insight into the potential efficacy of CroFab® against human envenomations by one of the most widely distributed rattlesnake species in North America.
Comparative proteomic analysis of venoms of neonate and adult Prairie Rattlesnake (Crotalus viridis viridis) from a discrete population in Colorado revealed a novel pattern of ontogenetic shifts in toxin composition for viperid snakes. The observed stage-dependent decrease of the relative content of disintegrins, catalytically active D49-PLA2s, L-amino acid oxidase, and SVMPs, and the concomitant increase of the relative abundance of paralytic small basic myotoxins and ohanin-like toxin, and hemostasis-disrupting serine proteinases, may represent an age-dependent strategy for securing prey and avoiding injury as the snake switches from small ectothermic prey and newborn rodents to larger endothermic prey. Such age-dependent shifts in venom composition may be relevant for antivenom efficacy and treatment of snakebite. However, applying a second-generation antivenomics approach, we show that CroFab®, developed against venom of three Crotalus and one Agkistrodon species, efficiently immunodepleted many, but not all, of the major compounds present in neonate and adult C. v. viridis venoms.
在此，我们描述并比较了新生和成年草原响尾蛇（绿响尾蛇）毒液的毒液组学和抗毒液组学特征。尽管新生和成年毒液都含有独特成分，但在蛋白质家族含量方面存在相似性。新生和成年毒液均由肌毒素、缓激肽增强肽（BPP）、磷脂酶A2（PLA2）、锌（2+）依赖性金属蛋白酶（SVMP）、丝氨酸蛋白酶、L-氨基酸氧化酶（LAAO）、富含半胱氨酸的分泌蛋白（CRISP）和解整合素家族组成。然而，观察到了定量差异，成年毒液中含有显著更高浓度的非酶毒性化合物，而新生毒液中含有更高浓度的消化前酶蛋白，如SVMP。为了评估这种毒液差异在蛇咬伤和蛇咬伤治疗背景下的相关性，我们在体外测试了常用抗蛇毒血清CroFab®识别成年和新生毒液的功效。这种比较表明，抗蛇毒血清使新生和成年毒液中的许多主要蛋白质家族（SVMP、CRISP、PLA2、丝氨酸蛋白酶和LAAO）发生免疫耗竭，而肌毒素是绿响尾蛇毒液的主要毒性成分之一，除了许多小肽外，未被CroFab®有效耗竭。因此，这些结果提供了绿响尾蛇毒液中存在的毒液化合物的全面目录，并对CroFab®针对北美分布最广的响尾蛇物种之一对人类中毒的潜在功效提供了新的分子见解。
对来自科罗拉多州一个离散种群的新生和成年草原响尾蛇（绿响尾蛇）毒液进行的比较蛋白质组学分析揭示了蝰蛇毒素组成个体发育变化的新模式。观察到的解整合素、催化活性D49-PLA2、L-氨基酸氧化酶和SVMP相对含量的阶段依赖性降低，以及麻痹性小碱性肌毒素和类ohanin毒素以及破坏止血的丝氨酸蛋白酶相对丰度的相应增加，可能代表了一种年龄依赖性策略，随着蛇从小型变温猎物和新生啮齿动物转向大型恒温猎物，确保捕获猎物并避免受伤所需。毒液组成的这种年龄依赖性变化可能与抗蛇毒血清的功效和蛇咬伤治疗有关。然而，应用第二代抗毒液组学方法，我们表明，针对三种响尾蛇属和一种蝮蛇属物种的毒液开发的CroFab®有效地免疫耗竭了新生和成年绿响尾蛇毒液中存在的许多但不是所有主要化合物。
