GPIb is involved in platelet aggregation induced by mucetin, a snake C-type lectin protein from Chinese habu (Trimeresurus mucrosquamatus) venom

Mucetin (Trimeresurus mucrosquamatus venom activator, TMVA) is a potent platelet activator purified from Chinese habu (Trimeresurus mucrosquamatus) venom. It belongs to the snake venom heterodimeric C-type lectin family and exists in several multimeric forms. We now show that binding to platelet glycoprotein (GP) lb is involved in mucetin-induced platelet aggregation. Antibodies against GPIb as well as the GPIb-blocking C-type lectin echicetin inhibited mucetin-induced platelet aggregation. Binding of GPIb was confirmed by affinity chromatography and Western blotting. Antibodies against GPVI inhibited convulxin- but not mucetin-induced aggregation. Signalling by mucetin involved rapid tyrosine phosphorylation of a number of proteins including Syk, Src, LAT and PLCgamma2. Mucetininduced phosphorylation of the Fcgamma chain of platelet was greatly promoted by inhibition of alpha(llb)beta(3) by the peptidomimetic EMD 132338, suggesting that phosphatases downstream Of alpha(llb)beta(3) activation are involved in dephosphorylation of Fcgamma. Unlike other multimeric snake C-type lectins that act via GPIb and only agglutinate platelets, mucetin activates alpha(llb)beta(3). Inhibition Of alpha(llb)beta(3) strongly reduced the aggregation response to mucetin, indicating that activation Of alpha(llb)beta(3) and binding of fibrinogen are involved in mucetin-induced platelet aggregation. Apyrase and aspirin also inhibit platelet aggregation induced by mucetin, suggesting that ADP and thromboxaneA(2) are involved in autocrine feedback. Sequence and structural comparison with closely related members of this protein family point to features that may be responsible for the functional differences.

Prolonged cardiac allograft survival in presensitized rats after a high activity Yunnan-cobra venom factor therapy

Complement-dependent antibody-mediated acute humoral rejection is the major obstacle of clinical transplantation across ABO incompatibility and human leukocyte antigen presensitization. We previously demonstrated that Yunnan-cobra venom factor (Y-CVF) cou

Purification, characterization and cytokine release function of a novel Arg-49 phospholipase A(2) from the venom of Protobothrops mucrosquamatus

Group IIA phospholipase A(2) (PLA(2)) are major components in Viperidae/Crotalidae venom. In the present study, a novel PLA(2) named promutoxin with Arg at the site 49 has been purified from the venom of Protobothrops muerosquamatus by chromatography. It

N49 phospholipase A(2), a unique subgroup of snake venom group II phospholipase A(2)

A novel phospholipase A(2) (PLA(2)) with Asn at its site 49 was purified from the snake venom of Protobothrops mucrosquamatus by using SP-Sephadex C25, Superdex 75, Heparin-Sepharose (FF) and HPLC reverse-phage C-18 chromatography and designated as TM-N49

Potent histamine-releasing activity of atrahagin, a novel snake venom metalloproteinase

Poisonous snakebite wound is a popular disease worldwide. However, the pathogenesis remains unclear. In the present study, a novel metalloproteinase atrahagin in Chinese cobra (Naja atra) snake venom was purified, using heparin-sepharose followed by Super

The role of anti-non-Gal antibodies in the development of acute humoral xenograft rejection of hDAF transgenic porcine kidneys in baboons receiving anti-Gal antibody neutralization therapy

Background. The present study was undertaken to determine the role of preformed and induced anti-non-Gal antibodies in the rejection of hDAF pig-to-baboon kidney xenotransplants after anti-Gal antibody neutralization therapy. Methods. Seven baboons receiv

A unique group of inactive serine protease homologues from snake venom

A number of inactive serine protease homologues (SPHs), which have poorly understood functions, have been identified in invertebrates and vertebrates. Recently, several SPH transcripts have been reported from snake venom glands, which provide potential ne

L-amino acid oxidase from Naja atra venom activates and binds to human platelets

An L-amino acid oxidase (LAAO), NA-LAAO, was purified from the venom of Naja atra. Its N-terminal sequence shows great similarity with LAAOs from other snake venoms. NA-LAAO dose-dependently induced aggregation of washed human platelets. However, it had n

Purification and characterization of a new L-amino acid oxidase from Daboia russellii siamensis venom

A new L-amino acid oxidase (designated as DRS-LAAO) was purified from Daboia russellii siamensis venom by ion-exchange, gel filtration and affinity chromatographies. DRS-LAAO is a homodimeric enzyme with a molecular weight of 120.0 kDa as measured by size

一种新的烙铁头蛇毒肌肉毒素

用分子筛和快速蛋白质液相色谱(FPLC)从烙铁头(Trimeresurus mucrosquamatus)蛇毒中分离了一个新的碱性肌肉毒素,命名为TMPB。它的分子量为16 000,等电点为9.2。用蛋白质序列仪测定了其N端24个氨基酸残基,TMPB与其他两个从同种蛇毒中分离到的碱性磷酯酶A_(2)的同源性分别为41.7%和54.2%。TMPB没有明显的磷酯酶A_(2)水解活性,但它的肌肉毒性和血小板聚集抑制活性却极强,其肌肉毒性和血小板聚集抑制活性可被肝素所抑制。

一个高活性的孟加拉眼镜蛇毒抗补体因子

用离子交换层析和分子筛从云南南部产的孟加拉眼镜蛇蛇毒中分离到一个高活性的抗补体因子。它表现出较强的体内、体外抗补体活性,其抗补体活性的比活力为1 515u/mg。纯化的抗补体因子在聚丙烯酰胺凝胶电泳中呈现一条带。经SDS-PAGE,确定其全分子量为149kD。还原性SDS-PAGE表明,它由3条多肽链共价结合而成,3条多肽链分子量分别为65.4kD、52.1kD和35.5kD。最小的一条多肽链在还原条件下呈现多态性,一般可见两条带(35.5kD和33.7kD)。过碘酸席夫试剂染色表明,其3条多肽链均含有糖。定量测定表明中性糖含量为1.78%,唾液酸含量为0.38%。其等电点为6.2。对其氨基酸组成分析表明它含有较多的酸性氨基酸。对其3条多肽链的N末端氨基酸序列进行了测定。

四个烙铁头蛇毒凝集素样蛋白基因的克隆与序列分析

从烙铁头蛇（Trimeresurus mucrosquamatus）的毒腺中提取mRNA，利用RT-PCR进行体外扩增，获得凝集素样蛋白基因，克隆至PMD18-T载体中，筛选出4种凝集素样蛋白基因（命名为TML-1、TML-2、TML-3和TML-4）。由基因序列推导出的氨基酸序列表明：TML-1，2，3，4序列中均有CRD结构。序列同源性比较和Cys位点分析推测：TML-1和TML-2可能分别是类似于flavocetin-A的蛇毒凝集素样蛋白的#alpha#亚基和#beta#亚基；TML-3可能类似于GPIb-bp的蛇毒凝集素样蛋白的#alpha#亚基，TML-4则可能是类似于IX/X-bp的蛇毒凝集素样蛋白的β亚基。

湖南烙铁头蛇毒中一个新的舒缓激肤增强肤( TmF)

通过70%冷甲醇抽提、Sephadex C i-15分子筛和反相高效液相色谱G8层析，从湖南产烙铁头蛇毒(Trineresurus muqua nwtus)冻干粉中纯化得到一个新的舒缓激肤增强肤(BPP)，命名为TmF。该小肤的氨基酸序列为p(irr (iy Arg Pro, Leti (iy Pro, Pro, Ile- Pro, Pro ( pau表示焦谷氨酸)。序列结果分析表明，TmF和已经分离得到的BPPs有很高的序列同源性。MSI- MS 测定其分子量为1 .1107 kD o TmF的生物学活性和药理学活性检测的结果表明，它增强舒缓激肤(BK)(1 mg/L)诱导的离体豚 鼠回肠纵行肌收缩的活性为(1 .13士0 .3)单位(mg/ L) ; TmF (5 .0 x 10- 0 mg/ kg)可以增强约(14士2) mmHg的由BK(5 . Ox 10-' mg/ kg)诱导的舒张压下降;在抑制剂试验中，不同剂量的TmF和5x1。一zmg的血管紧张素转化酶保温30 min，结果表明大约 2.0x10一3mg的TmF表现出对ACE水解活性的半数抑制率(IQo )。

不同地区舟山种和孟加拉种眼镜蛇的神经毒素

先前的工作发现，眼镜蛇孟加拉种的神经毒分布存在明显的地理差异，但是导致这种现象的原因还不是很清楚。该工作首先采集夏季的眼镜蛇种，进行分类学的鉴定，以排除因为种属分类、季节和年龄而导致的不确定因素对蛇毒成分产生影响。再从鉴定好的成年浙江舟山种和云南孟加拉种眼镜蛇蛇毒中分离获得短链神经毒素;同时采用RT PCR方法得到相应 的基因。结果从浙江舟山种眼镜蛇蛇毒中分离到两个神经毒素;从云南孟加拉种蛇毒中得到三个神经毒素。这些神经毒素都是短链神经毒素。说明该两个蛇种与已经报道的台湾及浙江的舟山种眼镜蛇很相似，蛇毒中都以短链神经毒素为主。另外泰国孟加拉种眼镜蛇蛇毒中则以长链神经毒素为主。这种现象很难用种群发生的前后或者地理距离的长短来解释，而可能与地区间的气候、地理环境或者眼镜蛇的食物差别相关。实验统计表明在云南、浙江和台湾这些条件基本相似，而泰国就有很大的不同。