Autophagy and other vacuolar protein degradation mechanisms

Autophagic degradation of cytoplasm (including protein, RNA etc.) is a non-selective bulk process, as indicated by ultrastructural evidence and by the similarity in autophagic sequestration rates of various cytosolic enzymes with different half-lives. The initial autophagic sequestration step, performed by a poorly-characterized organelle called a phagophore, is subject tofeedback inhibition by purines and amino acids, the effect of the latter being potentiated by insulin and antagonized by glucagon. Epinephrine and other adrenergic agonists inhibit autophagic sequestration through a prazosin-sensitive ₁-adrenergic mechanism. The sequestration is also inhibited by cAMP and by protein phosphorylation as indicated by the effects of cyclic nucleotide analogues, phosphodiesterase inhibitors and okadaic acid.Asparagine specifically inhibits autophagic-lysosomal fusion without having any significant effects on autophagic sequestration, on intralysosomal degradation or on the endocytic pathway. Autophaged material that accumulates in prelysosomal vacuoles in the presence of asparagine is accessible to endocytosed enzymes, revealing the existence of an amphifunctional organelle, the amphisome. Evidence from several cell types suggests that endocytosis may be coupled to autophagy to a variable extent, and that the amphisome may play a central role as a collecting station for material destined for lysosomal degradation.Protein degradation can also take place in a salvage compartment closely associated with the endoplasmic reticulum (ER). In this compartment unassembled protein chains are degraded by uncharacterized proteinases, while resident proteins roturn to the ER and assembled secretory and membrane proteins proceed through the Golgi apparatus. In thetrans-Golgi network some proteins are proteolytically processed by Ca²⁺-dependent proteinases; furthermore, this compartment sorts proteins to lysosomes, various membrane domains, endosomes or secretory vesicles/granules. Processing of both endogenous and exogenous proteins can occurr in endosomes, which may play a particularly important role in antigen processing and presentation. Proteins in endosomes or secretory compartments can either be exocytosed, or channeled to lysosomes for degradation. The switch mechanisms which decide between these options are subject to bioregulation by external agents (hormones and growth factors), and may play an important role in the control of protein uptake and secretion.     

小米乳在冰淇淋中的应用研究

详细介绍了以小米替代淀粉,研制的具有天然色香味的小米冰淇淋的生产工艺及配方。小米冰淇淋营养丰富、成本低、工艺简单、具有一定的推广价值。     

G蛋白偶联受体研究进展

G蛋白偶联受体（GPCRs）是体内最大的蛋白质超家族，根据结构的同源性，主要分为A、B、C3族．GPCRs配体的多样性决定配体结合域的多样性．受体分子内相互作用力的破坏、质子化、构象改变、与G蛋白的偶联及受体二聚化参与了GPCRs的活化过程，近年发现GPCRs的失敏和内吞对受体功能调节亦非常重要，本文拟综述以上内容的研究进展．     

乐清产缢蛏(Sinonovacula constricta)肉营养成分的研究

就缢蛏(Sinonovacula constricta)的营养成分进行了分析。用干燥法、凯氏定氮法、灼烧法、酸水解法、蒽酮比色法等方法,分别对其水分、蛋白质、灰分、脂肪、碳水化合物等营养成分作了测定。结果显示:水分、蛋白质、灰分、脂肪、碳水化合物占缢蛏肉百分比分别为85.56%,10.16%,2.82%,0.55%,0.93%。经比较,本实验缢蛏中的蛋白质成分高于其它地区。说明乐清产缢蛏是一种高蛋白、低脂肪,富含灰分的海味佳品。     

蛋白质芯片技术的研究进展

蛋白质芯片技术是生物化学和分子生物学上具有较大作用的生物检测技术。该文简要综述了该技术的发展概况、基本原理及目前应用，并指出了存在的问题和发展前景。     

人组织型纤溶酶原激活剂（t—PA）在大肠杆菌中的表达、复性及分离纯化

人组织型纤溶酶原激活剂（ｔ—ＰＡ）。ＤＮＡ重组在表达质粒ｐＤＲ７２０中，在Ｅ．ｃｏｌｉＣ６００中获得了表达，表达水平为２％．以醋酸钟显色ＰＡＧＥ凝胶回收ｔ—ＰＡ表达条带，进行复性处理．Ｒ性产物经Ｂｅｎｚａｍｉｄｉｎｅ—Ｓｅｐｈａｒｏｓｅ４Ｂ，Ｌｙｓｉｎｅ—Ｓｅｐｈａｒｏｓｅ４Ｂ亲和层析，纯化得到ＳＤＳ—ＰＡＧＥ银染一条带纯度，比活为为４，０００ｍｏｌ／ｓ·ｇ的人ｔ—ＰＡ．     

β－胡萝卜素在盐生杜氏藻（Dunaliella Salina）中积累机理的研究

以不同光照条件下生长的盐生杜氏藻为材料，通过测定叶绿体基质蛋白的含量发现，在低光照下叶绿体基质蛋白的含量为２６．４０ｇ／Ｌ，而在强光照下叶绿体基质蛋白为１１．５０ｇ／Ｌ。对这两种蛋白进行平板电泳皆为一条带，而在ＳＤＳ－凝胶电泳，低光照下为五条带（五个亚基），其Ｒ＿（ｆ１）值分别为０．４０，０．５０，０．６０，０；７０，０．７８；强光照下有四条带（四个亚基），其Ｒ＿（ｆ２）分别为０．５０，０．５８，０．６８，０．８０。通过改变ｐＨ值、提高温度和紫外线照射等，发现该蛋白具有酶活性并与β－胡萝卜素的合成有关。     

人C-反应性蛋白的提纯及检测

以胸腔渗出液为来源,用硫酸铵分级沉淀,DEAE—cellulose离子交换柱层析和快速蛋白质液相分离仪纯化了C—反应性蛋白。纯化的C—反应性蛋白经SDS—聚丙烯酰胺凝胶电泳、免疫电泳、Ouchterlony免疫扩散及快速蛋白质液相分离鉴定,纯度达100％。用这种纯化的C-反应性蛋白(CRP)免疫山羊所得的抗-CRP抗血清特异性好,用抗血清制就的CRP检测板和日本(KW)CRP检测板作对照测试,经060例验证,作Radit分析,P>0.05,表明两者质量无显著性差异。     

蛋白质微阵列技术及其影响因素的研究进展

简要介绍了蛋白质微阵列技术的基本原理和制备技术。重点讨论了蛋白质微阵列在制备和检测过程中的主要影响因素,如载体及其表面处理、探针、检测方法、制备技术等。指出蛋白质微阵列是一种极具发展前途的新型研究工具,在我国刚刚起步,需要加大投入和合理规划。     

Protein aggregation as primary and characteristic cell reaction to various stresses

Ehrlich carcinoma and EL-4 thymoma ascites cells were subjected in vitro to heat shock, ATP depletion, oxidative stress, Ca²⁺ overlading and iodoacetamide treatment. After the transient stresses, Triton (X-100)-insoluble TIS) fractions were isolated from the cells and analysed by electrophoresis and immunoblotting. All stresses used caused rapid aggregation of cell proteins. This was manifested in a signficant rise in protein content in the TIS fractions. The protein increase was mostly due to and increase in the insolubility of actin, 57 kDa protein of intermediate filaments, 70 kDa heat shock protein (HSP 70), and some specific proteins whose insolubilization was a characteristic sign for each type of cell injury. Different survival rates in the cell lines after either stress corrlated well with differences in their TIS protein accretion. Possible mechanisms for stress-induced protein aggregation and its relationship with cell viability are suggested.