Do antibody binding techniques identify polysomes synthesizing a specific protein?

Specific binding of antibody directed against MOPC-21 myeloma protein to MOPC-21 polysomes has been demonstrated. It was also shown that this same antibody bound specifically to the ribosomal subunits of these polysomes, suggesting that the antibody is not binding exclusively to the nascent polypeptide chains on the polysomes. The antibody was bound specifically to polysomes of a nonproducing variant XC1 which had been isolated in the presence of MOPC-21 myeloma protein while the antibody was not bound to XC1 polysomes isolated in the absence of myeloma protein. This suggests that the myeloma protein is adsorbed to the polysomes during the isolation procedure.     

In-depth Profiling and Quantification of the Lysine Acetylome in Hepatocellular Carcinoma with a Trapped Ion Mobility Mass Spectrometer

Hepatocellular carcinoma (HCC) is the third most common cause of cancer-related death worldwide with limited therapeutic options. Comprehensive investigation of protein posttranslational modifications in HCC is still limited. Lysine acetylation is one of the most common types of posttranslational modification involved in many cellular processes and plays crucial roles in the regulation of cancer. In this study, we analyzed the proteome and K-acetylome in eight pairs of HCC tumors and normal adjacent tissues using a timsTOF Pro instrument. As a result, we identified 9219 K-acetylation sites in 2625 proteins, of which 1003 sites exhibited differential acetylation levels between tumors and normal adjacent tissues. Interestingly, many novel tumor-specific K-acetylation sites were characterized, for example, filamin A (K865), filamin B (K697), and cofilin (K19), suggesting altered activities of these cytoskeleton-modulating molecules, which may contribute to tumor metastasis. In addition, we observed an overall suppression of protein K-acetylation in HCC tumors, especially for enzymes from various metabolic pathways, for example, glycolysis, tricarboxylic acid cycle, and fatty acid metabolism. Moreover, the expression of deacetylase sirtuin 2 (SIRT2) was upregulated in HCC tumors, and its role of deacetylation in HCC cells was further explored by examining the impact of SIRT2 overexpression on the proteome and K-acetylome in Huh7 HCC cells. SIRT2 overexpression reduced K-acetylation of proteins involved in a wide range of cellular processes, including energy metabolism. Furthermore, cellular assays showed that overexpression of SIRT2 in HCC cells inhibited both glycolysis and oxidative phosphorylation. Taken together, our findings provide valuable information to better understand the roles of K-acetylation in HCC and to treat this disease by correcting the aberrant acetylation patterns.     

Aberrant,ectopic expression of VEGF and VEGF receptors 1 and 2 in malignant colonic epithelial cells. Implications for these cells growth via an autocrine mechanism

Vascular endothelial growth factor A (referred to as VEGF) is implicated in colon cancer growth. Currently, the main accepted mechanism by which VEGF promotes colon cancer growth is via the stimulation of angiogenesis, which was originally postulated by late Judah Folkman. However, the cellular source of VEGF in colon cancer tissue; and, the expression of VEGF and its receptors VEGF-R1 and VEGF-R2 in colon cancer cells are not fully known and are subjects of controversy.     

探讨屎肠球菌的VanA调控人正常结直肠黏膜细胞FHC凋亡的机制

目的 探讨屎肠球菌的万古霉素替考拉宁A型抗性蛋白/D-丙氨酸-D-丙氨酸连接酶（Vancomycin Teicoplanin A-type resistance protein D-alanine-D-alanine ligase,vanA）调控人正常结直肠黏膜细胞FHC凋亡的机制。方法 在人正常结直肠黏膜细胞FHC中使用屎肠球菌感染,Annxin-V染色检测细胞凋亡情况。使用屎肠球菌的VanA蛋白刺激,检测FHC细胞凋亡情况、ROS水平以及ROS标志蛋白MDA、GSH和SOD的表达水平。ROS抑制Acetylcysteine处理VanA刺激的FHC细胞后,检测细胞凋亡相关蛋白的表达水平。结果 屎肠球菌与人正常结直肠黏膜细胞FHC共培养后,人正常结直肠黏膜细胞FHC的凋亡水平明显升高（t=2.876,P=0.045 2）,并且VanA蛋白能促进FHC凋亡水平（t=5.579,P=0.005 1）,同时细胞凋亡相关蛋白CLEAVED-CAS9、BAK的表达量上升,BCL-2的表达量下降。屎肠球菌的VanA蛋白刺激后,发现正常结直肠黏膜细胞FHC的ROS水平上升（t=10.190,P=0.000 5）,ROS标志蛋白MDA（t=4.315,P=0.012 5）和SOD（t=5.751,P=0.004 5）的表达水平上升,GSH（t=5.225,P=0.006 4）的表达水平下降,但是,ROS抑制剂Acetylcysteine能够抑制这种现象。结论 屎肠球菌的VanA通过提高细胞内ROS水平来促进人正常结直肠黏膜细胞FHC凋亡。     

Mark‐resight abundance and survival estimation of Indo‐Pacific bottlenose dolphins,Tursiops aduncus,in the Swatch‐of‐No‐Ground,Bangladesh

A mark‐resight analysis under Pollock's robust design was applied to Indo‐Pacific bottlenose dolphins Tursiops aduncus in the Swatch‐of‐No‐Ground (SoNG) submarine canyon, Bangladesh, during the winter seasons of 2005–2009. Information from sightings of photo‐identified individuals (1,144) and unmarked individuals generated abundance estimates of 1,701 (95% confidence interval [CI]= 1,533–1,888), 1,927 (95% CI = 1,851–2,006), 2,150 (95% CI = 1,906–2,425), and 2,239 (95% CI = 1,985–2,524) individuals for seasons 1–4, respectively. This makes the population among the largest assessed of the species. Overall apparent survival was estimated as 0.958 (95% CI = 0.802–0.992). Interseasonal probabilities of transitioning to an unobservable state were estimated as 0.045, 0.363, and 0.300 for years 1–2, 2–3, and 3–4, respectively, and the overall probability of remaining in an unobservable state was 0.688. These probabilities, together with an apparent increase in abundance during the study period, indicate that the identified dolphins are part of a larger superpopulation moving throughout a more extensive geographic area. Of the photo‐identified dolphins, 28.2% exhibited injuries related to entanglements with fishing gear. This implies a strong potential for fatal interactions that could jeopardize the conservation status of the population, which otherwise appears favorable.     

Radioimmunoassay of polypeptide hormones using immunochemically coated plastic tubes

A method has been developed for immobilisation of antisera on fresh plastic tubes through an immunochemical bridge. This type of immobilisation has been shown to be more consistent than direct adsorption on plastic. Such immunochemically coated antisera on plastic tube has been used in the development of a noncentrifugation radioimmunoassay. This assay system has been found to be technically as sound as the conventional method.     

Hemoglobin Dallas (α97(G4)Asn→Lys):functional characterization of a high oxygen affinity natural mutant

Hemoglobin Dallas, an α-chain variant with a substitution of lysine for asparagine at position 97(G4), was found to have increased oxygen affinity ($\text{p}{}_{\mathrm{<mtext>1</mtext><mtext>2</mtext>}}\text{= 1}\text{mmHg at pH}\text{7.3}\text{and}\text{20°C), diminished cooperativity (0n,}\text{the Hill coefficient}\text{= 1.7}$) and reduced Bohr effect (about 50%). Addition of allosteric effectors (such as 2,3-diphosphoglycerate, inositol hexakisphosphate and bezafibrate) led to a decrease in oxygen affinity and increase in cooperative energy. Kinetic studies at pH 7.0 and 20°C revealed that (i), the overall rate of oxygen dissociation is 1.4-fold slower than that for HbA and (ii), the carbon monoxide dissociation rate is unaffected. The abnormal properties of this hemoglobin variant can be atttributed to a more ‘relaxed’ T-state.     

Con A affinity glycoproteomics of normal human liver tissue

In order to establish the novel high throughput, high efficiency and Iow cost technological platform for the research of N-glycoproteomics, to resolve the significance of characteristic expression profile of glycoprotein and to find the proteins with biological functional importance, the glycoproteins with high-mannose core and the two antennary types were purified and enriched by the Con A affinity chromatography. Con A affinity protein expression profiles of normal human liver tissue were generated by using SDS-PAGE, two-dimensional electrophoresis (2-DE) followed by fast fluorescence staining based on multiplexed proteomics (MP) technology. 301 visible protein spots on the gel were detected and 85 of glycoproteins were further successfully identified via peptide mass fingerprinting (PMF) by a matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS/MS) and annotated to IPI databases. Identified glycoproteins definitely take part in the regulation of cell cycle and metabolic processes. The glycosylation sites were predicted with NetNGlyc 1.0 and NetOGlyc 3.1 software, meanwhile they were classified according to the geneontology methods. The construction of Con A affinity glycoprotein database of normal human liver tissue would contribute to the subsequent research.     

正常人和SLE患者血清中抗DNA抗体的研究

用亲和层析法对6例正常人和4例SLE患者血清中抗DNA抗体进行提取和定量研究,发现正常人血清中抗DNA抗体的组成IgM/IgG大于1,是以IgM为主的抗体,SLE患者血清中抗DNA抗体含量高,IgM/IgG小于1,是以IgG为主的抗体。用胰蛋白酶降解提取的抗DNA抗体再借助于SDS—聚丙烯酰胺凝胶电泳对正常人和SLE患者抗DNA抗体的结构进行初步探讨,电泳结果表明正常人和SLE患者纯化抗DNA抗体经胰蛋白酶降解以后,正常人在52.2Kd区有一特异性降解片段,而SLE患者则在33.6Kd区有明显的降解片段富集,且表明它们是抗DNA—IgG所产生的。这说明SLE患者血清中抗DNA—IgG不仅在数量上比正常人有所增加,而且在结构上也有所不同。此外,这两种不同的抗DNA—IgG被胰蛋白酶降解的速度也有差异。     

The finite number of global motion patterns available to symmetric protein complexes

In PDB, more than half of the entries are structure complexes and of these complexes, most are symmetric, composed of identical subunits. Complex formation is the way through which larger structures and even molecular machines are assembled and built in nature. In this work, we apply group theory and carry out a comprehensive study of the global motion patterns of protein complexes of various symmetries. The work presents for the first time a comprehensive list of all the symmetric, aesthetically pleasing, global motion patterns available to complexes of cyclic, dihedral, tetrahedral, or octahedral symmetry. Our results clearly demonstrate that complexes with the same symmetry will have the same global motion patterns and thus may function in a similar way, and that there are only a finite number of global motion patterns available to symmetric complexes as the number of protein symmetry groups is effectively finite. The work complements our current understanding of the principle of complex formation that has been mostly structure‐based by providing novel dynamics‐based insights. Furthermore, as dynamics is closely tied to function, these motion patterns can provide global insights into the general functional mechanisms of protein complexes.