Ramírez A N, Martin B M, Gurrola G B, Possani L D
Departamento de Bioquímica, Instituto de Biotecnología/UNAM, Cuernavaca, Morelos, México.
Toxicon. 1994 Apr;32(4):479-90. doi: 10.1016/0041-0101(94)90300-x.
A novel peptide, toxic to mice, was purified from the venom of the Mexican scorpion Centruroides limpidus limpidus, by means of gel filtration and ion exchange chromatography, followed by high performance liquid chromatography (HPLC). The complete amino acid sequence was determined by automatic Edman degradation of reduced and alkylated toxin, and by overlapping sequences of fragments of the toxin, generated by cleavage with proteinase V8 separated by HPLC. This toxin is composed of 66 amino acid residues, contains eight half-cystine residues, and is highly similar (91%) to the amino acid sequence deduced for toxin 1 of C. limpidus tecomanus and toxin 4 from C. noxius venom (89%). This peptide displaces the binding of radiolabeled toxin 2 of C. noxius from synaptosomal membranes of rat brain with superimposable kinetics, supporting the conclusion that it belongs to the beta-scorpion toxin class. Further characterization of C. l. limpidus toxin 1, as we have named it, was performed by means of competition experiments with monoclonal antibodies and various purified scorpion toxins, using an ELISA assay. A panel of six distinct monoclonal antibodies (mAB) against toxin 2 and 3 of C. noxius was used. From these, only three clones, originally named BCF1, BCF8 and BCF9, were able to recognize toxin 1 from C. l. limpidus.
从墨西哥蝎Centruroides limpidus limpidus的毒液中，通过凝胶过滤和离子交换色谱法，随后进行高效液相色谱（HPLC），纯化出一种对小鼠有毒的新型肽。通过对还原和烷基化毒素进行自动Edman降解，以及对通过HPLC分离的蛋白酶V8切割产生的毒素片段的重叠序列，确定了完整的氨基酸序列。这种毒素由66个氨基酸残基组成，含有8个半胱氨酸残基，与Centruroides limpidus tecomanus的毒素1和Centruroides noxius毒液中的毒素4推导的氨基酸序列高度相似（91%）。该肽以可叠加的动力学从大鼠脑突触体膜上取代放射性标记的Centruroides noxius毒素2的结合，支持其属于β-蝎毒素类别的结论。我们将其命名为Centruroides l. limpidus毒素1，通过使用ELISA测定法，与单克隆抗体和各种纯化的蝎毒素进行竞争实验，对其进行了进一步表征。使用了一组针对Centruroides noxius毒素2和3的六种不同单克隆抗体（mAB）。其中，只有三个最初命名为BCF1、BCF8和BCF9的克隆能够识别Centruroides l. limpidus的毒素1。
