Expression and characterization of the tau subunit of phosphorylase kinase

A cDNA encoding the entire tau subunit of rabbit skeletal muscle phosphorylase kinase was reconstructed and inserted into a plasmid containing the Escherichia coli ptac promoter and a constructed plasmid containing the ptac promoter and bacterial chloramphenicol acetyl transferase (CAT) gene, respectively. A significant phosphorylase kinase activity was found, in the first case. In the second case, a fused protein containing 73 amino acids from the CAT protein was obtained. After renaturation, the CAT-tau subunit protein shows enzymatic activity similar to the HPLC-purified and renatured tau subunit.     

Liver mitochondrial respiratory functions decline with age

Human liver mitochondrial respiration rates in Chinese populations of various ages were assayed with an oxygraph. In this study, State 3 and State 4 respiration rates, respiratory control ratio (RCR), and ADP/O ratio were measured for 35 Chinese subjects of 31 to 76 years old. We found a significant negative correlation between age and respiratory control and ADP/O ratios tested. Moreover, the respiratory control and ADP/O ratios decreased with the increase of age. These findings suggest that a substantial fall in mitochondrial oxidative capacity in ageing liver may be an important contributor to the ageing process.     

Expression of lipoprotein lipase gene in combined lipase deficiency

The expression of the gene for lipoprotein lipase (LPL) was studied in brown adipose tissue and the liver of combined lipase deficient (cld/cld) and unaffected mice. The mRNA specific for LPL was detected in both animals. Although the size of LPL mRNA in cld mice was similar to that of unaffected mice, the mRNA concentration in affected animals was higher than in unaffected animals. We also studied the LPL gene mutation in cld mice by Southern blot analysis. No restriction fragment length polymorphisms were observed after digestion with 16 endonucleases. These data indicate that there is no gene insertion or deletion, but do not exclude the possibility of point mutation in the LPL structural gene. However, the present results agree with the hypothesis that the genetic defect in cld is not due to a mutation in the LPL structural gene, but instead involves the defective post-translational processing of LPL or defective cellular function affecting transport and secretion of this enzyme group.     

Two LINE 1 repeats in rat

One LINE 1 repeat has been located 661 bp downstream from the last albumin exon and another approx. 10 kbp downstream from the last alpha-fetoprotein exon in the rat genomic DNA. The LINE 1 repeat following the albumin gene is truncated at its 5' end and is 1204 nucleotides long. The 5' end of the longer repeat downstream from the alpha-fetoprotein gene has not been determined. The two repeats have 95% homology with each other, with the exception of a short diverse 3' end sequence just preceding the putative polyadenylation signal.     

Studies of blocking mechanisms of carbachol-induced polyphosphoinositide turnover in rat cortical synaptosomes by neuroactive drugs

1. The incorporation of 32Pi into 4 phospholipids of rat cortical synaptosomes was altered in the presence of carbachol (1 mM), viz. a decrease of phosphatidylinositol-4,5-bisphosphate and phosphatidyl-4-phosphate by 34 and 21%, and an increase of phosphatidylinositol and phosphatidic acid by 52 and 96% of basal controls respectively. 2. The IC30 values calculated from the dose-response curves for drugs affecting carbachol-induced 32Pi incorporation into these phospholipids, and [3H]QNB binding to the cortical synaptosomes were similar for the typical antimuscarinic agents (i.e. atropine, pirenzepine and trihexyphenidyl), and tricyclic antidepressants (i.e. amitriptyline, doxepin and imipramine) studied. 3. The IC30 values obtained for drugs affecting carbachol-induced 32Pi incorporation into these phospholipids, and high potassium-induced 45Ca2+-uptake by this preparation were similar for neuroselective calcium channel blockers (i.e. cinnarizine and flunarizine) studied. 4. Our results suggest that the neuroactive drugs studied can either act at, or beyond the receptor level, perhaps on the availability of calcium ion, to block carbachol-induced polyphosphoinositide turnover in rat cortical synaptosomes.     

小麦间期集缩染色质的超微结构研究

本文以普通小麦(Triticum aestivum L.)根端分生组织为材料,在透射电镜下对间期细胞核内的集缩染色质的高层次结构进行了研究。在其中观察到直径约为20—25nm、50nm及110—120nm 的不同等级染色线,并且发现直径110—120nm 的染色线是由50nm 的染色线组成的,而直径约50nm 的染色线是由20—25nm 的染色线组成的。对这三个层次染色质结构之间的集缩方式进行了讨论。     

The use of micropreparative electrophoresis of protein/peptide isolations for primary structure determinations

High-performance electrophoresis chromatography (HPEC) is a recent development that features continuous-elution gel electrophoresis for isolating proteins or peptides in range of 1 to 300 microgram quantities. Column gel electrophoresis is conducted under thermostated conditions, and the field voltage can be varied within a run with a programmable power supply. Applications of this apparatus in protein purification are presented to demonstrate the utility of the (Model 230A) HPEC. These examples include on-line detection with direct analyte recovery of highly purified sample, which mimics high-performance liquid chromatography, for subsequent structure-function characterization. A method to remove salts from sodium dodecyl sulfate electrophoresed samples for subsequent sequencing or amino acid analysis is described. This desalting procedure recovers from 90%-95% of the sample and employs a low molecular weight cut-off membrane during sample centrifugation onto a polyvinylidene difluoride membrane. Subsequent washings are performed to efficiently remove salts, free amino acids and detergents that are known to interfere with sequence analysis. Sequence information such as initial recovery, repetitive yields and chromatogram comparisons are presented to demonstrate the utility of this procedure when used following isolation of sample with HPEC.     

植物单盐毒害和离子拮抗机理研究Ⅰ.单盐和混合盐对植物原生质体膜某些特性、超氧物歧化酶和过氧化氢酶活性的影响

单盐(KCl, CaCl_2或MgCl_2)和混合盐(KC_1+CaCl_2或KCl+MgCl_2)对植物原生质体完整率、存活率和膜透性等均有明显影响。K~+、Ca~(2+)或Mg~(2+)等单种阳离子明显降低原生质体膜完整率和存活率而增加其物质渗漏量,其中以单价阳离子K~+的影响为甚。上述单种阳离子还明显降低小麦幼叶超氧物歧化酶(SOD)和过氧化氢酶活性。只有由单价和二价阳离子组成的平衡混合盐才能使原生质体维持较高的完整率、存活率和较正常的膜透性.并能使细胞维持较高的SOD和过氧化氢酶活性。 认为单盐毒害机理可能是首先引起细胞膜发生不正常的膜相变或细胞累积较多的有害氧自由基,引起膜脂发生过氧化或脱酯化而破坏膜结构。在离子平衡混合盐作用下,膜系才能维持正常液晶相,具有较高活性的SOD和过氧化氢酶等生物保护性酶系是离子拮抗作用之原因。