Interaction between the conserved region in the C-terminal domain of GRK2 and rhodopsin is necessary for GRK2 to catalyze receptor phosphorylation

The C-terminal domain of G protein-coupled receptor kinases (GRKs) consists of a conserved region and a variable region, and the variable region has been shown to direct the membrane translocation of cytosolic enzymes. The present work has revealed that the C-terminal domain may also be involved in kinase-receptor interaction that is primarily mediated by the conserved region. Truncation of the C-terminal domain or deletion of the conserved region in this domain of GRK2 resulted in a complete loss of its ability to phosphorylate rhodopsin and in an obvious decrease in its sensitivity to receptor-mediated phosphorylation of a peptide substrate. On the contrary, deletion of the betagamma subunit binding region in the C-terminal domain of GRK2 did not significantly alter the ability of the enzyme to phosphorylate rhodopsin. In addition, the recombinant proteins that represent the C-terminal domain and the conserved region of GRK2 could inhibit GRK2-mediated phosphorylation of rhodopsin and receptor-mediated activation of GRK2 but not GRK2-mediated phosphorylation of the peptide substrate. Furthermore, the conserved region as well as the C-terminal domain could directly bind rhodopsin in vitro. These results indicate that the C-terminal domain, or more precisely, the conserved region of this domain, is important for enzyme-receptor interaction and that this interaction is required for GRK2 to catalyze receptor phosphorylation.     

Huntingtin interacting protein 1 induces apoptosis via a novel caspase-dependent death effector domain

Huntington disease is a devastating neurodegenerative disease caused by the expansion of a polymorphic glutamine tract in huntingtin. The huntingtin interacting protein (HIP-1) was identified by its altered interaction with mutant huntingtin. However, the function of HIP-1 was not known. In this study, we identify HIP-1 as a proapoptotic protein. Overexpression of HIP-1 resulted in rapid caspase 3-dependent cell death. Bioinformatics analyses identified a novel domain in HIP-1 with homology to death effector domains (DEDs) present in proteins involved in apoptosis. Expression of the HIP-1 DED alone resulted in cell death indistinguishable from HIP-1, indicating that the DED is responsible for HIP-1 toxicity. Furthermore, substitution of a conserved hydrophobic phenylalanine residue within the HIP-1 DED at position 398 eliminated HIP-1 toxicity entirely. HIP-1 activity was found to be independent of the DED-containing caspase 8 but was significantly inhibited by the antiapoptotic protein Bcl-x(L), implicating the intrinsic pathway of apoptosis in HIP-1-induced cell death. Co-expression of a normal huntingtin fragment capable of binding HIP-1 significantly reduced cell death. Our data identify HIP-1 as a novel proapoptotic mediator and suggest that HIP-1 may be a molecular accomplice in the pathogenesis of Huntington disease.     

Tumor necrosis factor (TNF) receptor superfamily member TACI is a high affinity receptor for TNF family members APRIL and BLyS

An expression cloning approach was employed to identify the receptor for B-lymphocyte stimulator (BLyS) and identified the tumor necrosis factor receptor superfamily member TACI as a BLyS-binding protein. Expression of TACI in HEK293T cells confers on the cells the ability to bind BLyS with subnanomolar affinity. Furthermore, a TACI-Fc fusion protein recognizes both the cleaved, soluble form of BLyS as well as the membrane BLyS present on the cell surface of a recombinant cell line. TACI mRNA is found predominantly in B-cells and correlates with BLyS binding in a panel of B-cell lines. We also demonstrate that TACI interacts with nanomolar affinity with the BLyS-related tumor necrosis factor homologue APRIL for which no clear in vivo role has been described. BLyS and APRIL are capable of signaling through TACI to mediate NF-kappaB responses in HEK293 cells. We conclude that TACI is a receptor for BLyS and APRIL and discuss the implications for B-cell biology.     

Electrospray ionization/mass spectrometry compatible reversed-phase separation of phospholipids: piperidine as a post column modifier for negative ion detection

An electrospray ionization (ESI) compatible separation of phospholipids (PL), phosphatidylglycerol (PG), phosphatidylethanolamine (PE), and phosphatidylcholine (PC), was performed on a C18 column by reversed phase High Performance Liquid Chromatography (HPLC) with minimal ESI suppression. The mobile phase, used isocratically, consisted of methanol and water. ESI was used to efficiently transfer the ions present in solution to the gas phase for mass spectrometric (MS) detection. Formation of negative ions was reinforced by incorporating piperidine post column. Limits of detection (LOD) and limits of quantitation (LOQ) were experimentally determined to be 20 and 60 fmol/microl, respectively, when acquiring data in the selected ion monitoring (SIM) mode monitoring three ions with a single quadrupole MS. When acquiring data from m/z 110-900 in the scanning mode, the LOD and LOQ were experimentally determined to be 1 pmol/microl and 3 pmol/microl. When acquiring product ion spectra for m/z 747, the LOD and LOQ were experimentally determined to be 446 attomol/microl and 1.3 fmol/microl, respectively.     

基于胃含物和碳、氮稳定同位素研究浙江南部近海蓝圆鲹的摄食生态

蓝圆鲹是浙江南部近海的重要经济鱼类。本文根据2020年5月、8月、11月和2021年1月在浙江南部近海进行的底拖网调查,应用胃含物分析和碳氮稳定同位素分析对浙江南部近海蓝圆鲹的摄食习性进行研究。结果表明: 浙江南部近海蓝圆鲹δ¹³C平均值为(-16.55±0.60)‰,范围为-17.76‰～-15.25‰,与叉长呈显著负相关;蓝圆鲹δ¹⁵N平均值为(11.76±0.88)‰,范围为9.06‰～13.03‰,与叉长呈显著正相关。根据δ¹⁵N值计算浙江南部近海蓝圆鲹的平均营养级为3.89±0.26。胃含物分析表明,浙江南部近海蓝圆鲹的主要饵料类群为鱼类、虾类、蟹类、头足类、多毛类和小型甲壳类。稳定同位素分析表明,虾类对浙江南部近海蓝圆鲹的营养贡献率最高(40%～84%),其次为多毛类、小型甲壳类、蟹类、头足类和鱼类。蓝圆鲹的摄食习性有明显的生长变化,随着蓝圆鲹叉长的增加,其倾向于摄食更高营养级的饵料生物。     

A highly pathogenic strain of Staphylococcus sciuri caused fatal exudative epidermitis in piglets

Staphylococcus sciuri are important human pathogens responsible for endocarditis, peritonitis, septic shock, urinary tract infection, pelvic inflammatory disease and wound infections. However, little information is known regarding the pathogenicity of S. sciuri to animals. From the pericardial fluid of a diseased piglet with exudative epidermitis (EE), we isolated a strain of Staphylococcus in pure culture. Surprisingly, this isolate was a member of S. sciuri rather than S. hyicus as identified by its biochemical traits and also by analysis of 23S ribosomal DNA using Internal Transcribed Spacer PCR. In addition, inoculation of newborn piglets with 1x10(10) CFU of the isolate by oral feeding or intra-muscular injection successfully reproduced EE in piglets, which suggested that the oral intake of the pathogen by the animals is one of the major routes of exposure. These unexpected findings prioritized S. sciuri as important zoonotic agents, which may have ramifications for human medicine.     

甘蔗节间伸长过程赤霉素生物合成关键基因的表达及相关植物激素动态变化

以甘蔗(Saccharum officinarum)优良品种桂糖42号(GT42)为研究材料, 分别于未伸长期(9-10叶龄以前) (Ls1)、伸长初期(12-13叶龄) (Ls2)和伸长盛期(15-16叶龄) (Ls3)取甘蔗第2片真叶(自顶部起)对应的节间组织, 测定其赤霉素(GA)、生长素(IAA)、油菜素甾醇(BR)、细胞分裂素(CTK)、乙烯(ETH)和脱落酸(ABA)的含量, 并通过实时荧光定量PCR (qRT-PCR)分析赤霉素合成途径关键基因GA₂₀氧化酶基因(GA20-Oxidase1)、赤霉素受体基因(GID1)和DELLA蛋白编码基因(GAI)的差异表达。结果表明, 在甘蔗伸长期间, GA和IAA含量呈现上升趋势, CTK和ABA含量呈下降趋势, ETH含量先上升后下降, BR含量则变化不明显; GA20-Oxidase1和GID1的表达呈上升趋势, 而GAI的表达则呈下降趋势, 这与相关植物激素的变化基本一致。综上, 甘蔗节间伸长过程主要与GA和IAA相关, 其次为CTK和ABA, 而ETH受到IAA的调控影响节间伸长; 植物激素间通过相互作用调控GA20-Oxidase1、GID1和GAI的表达, 影响GA含量和GA的信号转导过程, 进而影响甘蔗节间的伸长。该研究揭示了甘蔗节间伸长过程中赤霉素生物合成途径和信号转导关键基因的差异表达及植物激素含量的动态变化规律。     

盆底超声评价在妊娠妇女盆底结构、功能以及预后上的价值

为探究盆底超声检查评估分析不同的分娩方式对女性晚期妊娠和产后盆底结构、功能及预后效果的影响,本研究选取收治的100例顺利生产的孕妇作为本实验研究对象,根据分娩方式的不同将其分为自然分娩组与剖宫产组,各组50例。在两组孕妇妊娠晚期和产后分别通过盆底超声检测盆底结构和功能(膀胱旋转角,尿道膀胱后角,肛提肌裂孔面积和肛提肌裂孔周长);以及在产后第6~8周和盆底康复治疗后进行压力性尿失禁发生率调查问卷表填写及护垫实验。研究发现,阴道自然分娩比选择性剖宫产对妊娠晚期和产后孕妇盆底结构和功能造成的损伤更为显著;盆底损伤较严重的孕妇压力性尿失禁程度同样较严重;而盆底康复治疗成功的压力性尿失禁患者,盆底超声检测发现孕妇产后盆底结构和功能得到有效改善。本研究结果说明,盆底超声评价在检测不同分娩方式对妊娠晚期和产后孕妇的盆底结构、功能及预后存在一定的预测和预防作用。     

右旋糖酐酶研究进展

右旋糖酐酶是一种将高分子右旋糖酐催化降解为低分子量多糖的水解酶。该酶及其催化产物在医药、食品、化工等工业领域具有重要的应用价值与广泛的工业用途,因此近年来国内外对右旋糖酐酶的研究逐渐增多。结合文献记载及本实验室研究成果,对右旋糖酐酶的研究进展及其工业应用进行综述,并对当前有关该酶研究的热点和重点、国内右旋糖酐酶研究存在的问题以及未来的研究趋势提出了见解。