Rajesh R, Nataraju A, Gowda C D R, Frey B M, Frey F J, Vishwanath B S
Department of Studies in Biochemistry, University of Mysore, Manasagangothri, Mysore 570006, India.
Biochimie. 2006 Oct;88(10):1313-22. doi: 10.1016/j.biochi.2006.06.007. Epub 2006 Jul 5.
Latex glycoprotein (LGP) from Synadenium grantii latex was purified by the combination of heat precipitation and gel permeation chromatography. LGP is a heat stable protein even at 80 degrees C showed a sharp single band both in SDS-PAGE as well as in native (acidic) PAGE. LGP is a monomeric protein appears as single band under reducing condition. It is a less hydrophobic protein showed sharp single peak in RP-HPLC with retention time of 13.3 m. The relative molecular mass of LGP is 34.4 kDa. CD spectrum of LGP explains less content of alpha-helix (7%), and high content of beta-pleated sheets (48%) and random coils (46%). The N-terminal sequence of LGP is D-F-P-S-D-W-Y-A-Y-E-G-Y-V-I-D-R-P-F-S. Purified LGP is a fibrinogen degrading protease hydrolyses all the three subunits in the order of Aalpha, Bbeta and gamma. The hydrolytic pattern is totally different from plasmin as well as thrombin. LGP reduces recalcification time from 165 to 30 s with citrated human plasma but did not show thrombin like as well as factor Xa-like activity. Although LGP induces procoagulant activity, it hydrolyses partially cross-linked fibrin clot. It hydrolyses all the subunits of partially cross-linked fibrin clot (alpha- chains, beta-chain and gamma-gamma dimer). LGP is a serine protease, inhibited by PMSF. Other serine protease inhibitors, aprotinin and leupeptin did not inhibit the caseinolytic activity as well as fibrinogenolytic activity. We report purification and characterization of a glycoprotein from Synadenium grantii latex with human fibrino(geno)lytic activity.
通过热沉淀和凝胶渗透色谱相结合的方法，从格氏腺叶胶乳中纯化出乳胶糖蛋白（LGP）。LGP是一种热稳定蛋白，即使在80摄氏度时，在SDS-PAGE以及天然（酸性）PAGE中均显示出清晰的单一条带。LGP是一种单体蛋白，在还原条件下呈现为单一条带。它是一种疏水性较低的蛋白，在反相高效液相色谱（RP-HPLC）中显示出尖锐的单峰，保留时间为13.3分钟。LGP的相对分子质量为34.4 kDa。LGP的圆二色光谱表明其α-螺旋含量较低（7%），β-折叠片层含量较高（48%），无规卷曲含量较高（46%）。LGP的N端序列为D-F-P-S-D-W-Y-A-Y-E-G-Y-V-I-D-R-P-F-S。纯化后的LGP是一种纤维蛋白原降解蛋白酶，可按Aα、Bβ和γ的顺序水解所有三个亚基。其水解模式与纤溶酶和凝血酶完全不同。LGP可将枸橼酸化人血浆的再钙化时间从165秒缩短至30秒，但未表现出凝血酶样以及因子Xa样活性。尽管LGP诱导促凝血活性，但它可部分水解交联纤维蛋白凝块。它可水解部分交联纤维蛋白凝块的所有亚基（α链、β链和γ-γ二聚体）。LGP是一种丝氨酸蛋白酶，被苯甲基磺酰氟（PMSF）抑制。其他丝氨酸蛋白酶抑制剂，如抑肽酶和亮抑肽酶，并未抑制酪蛋白水解活性以及纤维蛋白原水解活性。我们报道了从格氏腺叶胶乳中纯化和鉴定一种具有人纤维蛋白（原）溶解活性的糖蛋白。
