Oscillatory change of SR-protein kinase activities during oocyte maturation meiosis in fish

The SR-protein kinase activity was analyzed and the cytological changes were observed during oocyte maturation in bisexual transparent color crucian carp ( Carassius auratus color variety). The results revealed that the SR-protein kinase activity was sensitive to the artificially induced spawning hormones, and the change of oscillatory activity was similar to that of the maturation-promoting factor (MPF) kinase that regulates meiotic cell cycle in fish.     

Protein kinase C regulation of GABAA receptors

Pharmacological studies with drugs that activate or inhibit several protein kinase C (PKC) isozymes have identified the PKC family of serine-threonine kinases as important in the regulation of -aminobutyric acid type A (GABA_A) receptor function. PKC modulates GABA_A receptor surface density, chloride conductance and receptor sensitivity to positive allosteric modulators such as neurosteroids, ethanol, benzodiazepines and barbiturates. Recent studies using PKC isozyme-selective reagents and gene-targeted mice have begun to identify critical roles for three isozymes, PKCII, PKC and PKC, in various aspects of GABA_A receptor regulation. Progress in this field touches upon therapeutic areas that are of great clinical importance such as anxiety and addiction. Increased understanding of how PKC regulates GABA_A receptors and which PKC isozymes are involved holds promise for development of new treatments for diverse neuropsychiatric disorders.Received 2 August 2004; received after revision 17 August 2004; accepted 21 August 2004     

抗三尖杉酯碱的HL60细胞蛋白质磷酸化

研究了抗三尖杉酯碱的ＨＬ６０细胞蛋白质磷酸化的变化。经差速离心得到纯膜蛋白，抗性细胞有一高度磷酸化的１１０ｋｕ的蛋白质存在，免疫沉淀ｃ－ＫＡＦ－１蛋白激酶，抗性细胞内ｃ－ＲＡＦ－１蛋白激酶磷酸化程度明显提高，其活性被蛋白激酶Ｃ抑制剂ＣａｌｐｈｏｓｔｉｎＣ明显地抑制。结果表明：ＨＬ６０细胞对三尖杉酯碱的抗药性与蛋白质高度磷酸化有关；抗性细胞内ｃ－ＲＡＦ－１蛋白激酶磷酸化程度明显提高，可能与多药抗药性和抗细胞调亡有关。     

大肠杆菌中融合表达汉滩病毒S,M基因片段的研究

为在大肠杆菌中融合表达汉滩病毒囊膜糖蛋白G1与NP含主要抗原位点的片段 .将汉滩病毒 76 118株S基因编码区 5′端约 0 .7kb的片段与M基因编码G1的片段连接 ,克隆入 pGEX 4T2 ,构建嵌合基因原核表达载体 pGEX 4T2 S 0 .7G1,在大肠杆菌XL1 Blue中诱导GST S0 .7G1融合蛋白的表达 .表达产物用ELISA和Westernblot进行鉴定 .限制性内切酶酶切鉴定证明 ,成功构建了嵌合原核表达载体 pGEX 4T2 S0 .7G1.经IPTG诱导后 ,ELISA活性测定结果表明 ,该融合蛋白可与抗汉坦病毒NPmAb特异性结合 .Westernblot结果显示 ,诱导出相对分子质量 (Mr)大于 1× 10 5的S0 .7G1与GST的融合蛋白 ,并有一系列大小不等的可与抗汉坦病毒NPmAb特异性结合的蛋白带 .获得具有特异性结合活性的融合蛋白GST S0 .7G1,为汉滩病毒基因工程疫苗的研制奠定了基础     

从粉丝废水中提取蛋白质的研究

用生物化学方法从粉丝废水中提取蛋白质，其提取物粗蛋白含量高，氨基酸种类较全，能作为高蛋白饲料添加剂使用。     

新型Protein A免疫吸附膜材料制备及其特性

以纤维素复合膜为基质合成了Protein A免疫吸附材料，研究了微环境对固定化的Protein A吸附性能的影响，优化了合成方法并对Protein A免疫吸附膜材料的性能进行了评价。实验表明，当每g复合膜上的环氧基团为0．74mmol，间隔臂长度为18个原子，溶液pH为7－8时，Protein A免疫吸附膜能够达到良好的吸附性能和再生性能，每g膜对IgG的最大吸附能力为12．84mg，在连续使用100次时，仍能达到10．08mg。制备的Protein A免疫吸附膜柱具备良好的流动特性和抗压能力，同时非特异吸附小，能够满足免疫吸附治疗的要求。     

Spermine protects protein kinase C from phospholipid-induced inactivation

Phosphatidylserine (PS), an activator of protein kinase C (PKC) in the assay of protein phosphorylation, inhibited this enzyme in a time-dependent manner following preincubation in the absence of Ca²⁺. The phospholipid-induced inactivation of kinase activity was dependent on the PS content and on the charge density of liposomes. This inactivation of PKC could be reduced, but not completely eliminated, by addition of Ca²⁺. In the present work the effect of a naturally occurring polyamine (spermine) on the PS-induced inactivation of PKC was investigated. The presence of spermine during preincubation without Ca²⁺ was effective in suppressing the PS-induced inactivation of PKC over the period (20 min) required for PS to inhibit the enzyme by 95%. PKC exists in two membrane-bound states: a reversible one which can be dissociated by Ca²⁺ chelators (membrane-associated form) and an irreversible one which is chelator-stable (membrane-inserted form). Gel filtration experiments on the PKC-PS complex formed in the presence of Ca²⁺ indicated that less insertion of enzyme into liposomes occurred in the presence of spermine and that the kinase activity of the reversibly membrane-associated PKC was protected from PS inactivation.     

从猪血综合提取凝血酶等生化产品的研究

采用现代工业生物化学技术 ,对猪血进行了综合利用研究 ,从中提取了高价值的凝血酶、铜、锌 -超氧化物歧化酶和水解蛋白等三种生化产品以及副产品饲料血粉 ,从而使猪血整个变废为宝 ,并解决了因猪血废弃造成环境污染的问题。