High surface hydrophobicity ofStaphylococcus aureus as revealed by hydrophobic interaction chromatography

Hydrophobic interaction chromatography (HIC) on Octyl Sepharose^R in a column procedure was used to compare the relative surface hydrophobicity ofStaphylococcus aureus reference strains, protein A-negative mutants, and strains isolated from bovine mastitis. High protein A-producing strains (Cowan 1 and clinical isolate SA 17970) showed a higher relative surface hydrophobicity than did strains producing a low amount of protein A. One encapsulatedS. aureus strain (Smith diffuse) did not bind to the gel, while an unencapsulated variant showed binding properties similar to weak protein A-producing strains. Studies onS. aureus strains isolated from bovine mastitis revealed a good correlation between adsorption to Octyl Sepharose and the production of protein A. Results indicate that protein A and probably other surface proteins such as fibronectin-binding protein contribute to the high relative surface hydrophobicity ofS. aureus.     

Glycoprotein of protein bodies fromLupinus luteus L. seeds

Proteins of protein bodies from lupin cotyledons were subjected to affinity chromatography on a column of Con A-Sepharose. About 42% of them contained sugar component. Globulin fractions soluble and precipitated at pH 4.4 contained 77% and 66% of glycoprotein, respectively.     

Protein composition of nuclear 14 S ribonucleoprotein particles containing poly (A)

Nuclear 14 S RNP particles containing poly (A) from Ehrlich ascites carcinoma cells and rat liver were purified by re-sedimentation in sucrose gradients, by Cs₂SO₄ density gradient centrifugation and by affinity chromatography on a poly (dT)-Sepharose column. Proteins of these RNP particles were electrophoresed in urea and SDS-polyacrylamide gels. RNP particles of ascites carcinoma cells contain two main bands having molecular weights of 51 000 and 69 000 daltons, respectively, and two or three minor components.     

Complex-type carbohydrates of apolipoprotein-B of human plasma low-density lipoproteins

Two fractions of glycopeptides containingN-glycosidic asparagine-linked glycans were isolated by concanavalin-A-Sepharose affinity chromatography from Pronase digests of apolipoprotein-B of human low-density lipoproteins. Methylation analysis indicated that the non-binding fraction contains about 0.5 mol complex-type tri- or tetra-antennary oligosaccharides per mol lipoprotein. Structures containing a bisectingN-acetylglucosamine were also detected in this fraction. The weakly binding fraction from the chromatography contains the majority of the complex oligosaccharides of apolipoprotein-B, i.e. 5–6 mol of bi-antennary chains of the transferrin-type per mol of lipoprotein. In addition to sialylated structures, branches containing a terminalN-acetylglucosamine residue were detected on the complex-type glycans of apolipoprotein-B.     

Haemagglutinin inTriticale

Saline extracts of several varieties ofTriticale had haemagglutinin activity against rabbit, rat and fowl erythrocytes. In contrast to the wheat germ lectin theTriticale lectin was inactive against human B, 0 blood group type erythrocytes and rather high concentrations of the lectin are needed to agglutinate human A blood group type erythrocytes. TheTriticale lectin was purified about 20-fold with a 10% recovery of activity from one of the varieties (DTS 138) by (NH₄)₂SO₄ fractionation followed sequentially by chromatography on DEAE-cellulose and sulphopropyl-Sephadex. Approximately 4 μg of the purified lectin caused visible agglutination with trypsinised rabbit erythrocytes. Among a variety of sugars tested D-glucose, D-mannose and N-acetyl-D-glucosamine (2·5-7·5mM) caused inhibition of agglutination.     

Changes in non-cellulosic cell-wall polysaccharides during the growth of carrot cells in suspension cultures

The cell-wall composition of carrot (Daucus carota L.) cells has been studied during their growth in suspension culture. Pectic and hemicellulosic polymers were fractionated according to molecular size by a Sepharose 4B column. Polyuronides in the pectic fraction were resolved into high- and low-molecular-weight components. The low-molecular-weight polyuronides were relatively free of neutral sugars and showed a marked increase during the growth of the cell wall. Hemicellulosic polysaccharides were of disperse molecular size. As cell expansion proceeded, the contents of glucose and xylose in the high-molecular-weight region increased while those in the low-molecular-weight fraction decreased. Removal of auxin from the medium apparently caused degradation of high-molecular-weight polymers in both the pectic and hemicellulosic fractions.     

Distinction between cofactor-dependent and-independent phosphoglycerate mutases by chromatography on Cibacron Blue-Sepharose

The binding of phosphoglycerate mutases from a variety of sources to Cibacron Blue-Sepharose has been examined. Those enzymes which are dependent on 2,3-bisphosphoglycerate (BPG) for activity bind to the immobilized dye and can be eluted by BPG. Those enzymes which are independent of BPG do not bind to the immobilized dye. The possible structural signifi-cance of this distinction is discussed.     

Identification of nucleolus-localized PTEN and its function in regulating ribosome biogenesis

The tumor suppressor PTEN is a lipid phosphatase that is found mutated in different types of human cancers. PTEN suppresses cell proliferation by inhibiting the PI3K-Akt signaling pathway at the cell membrane. However, PTEN is also demonstrated to localize in the cell nucleus where it exhibits tumor suppressive activity via a different, unknown mechanism. In this study we report that PTEN also localizes to the nucleolus and that nucleolar PTEN plays an important role in regulating nucleolar homeostasis and maintaining nucleolar morphology. Overexpression of nuclear PTEN in PTEN null cells inhibits Akt phosphorylation and reduces cell size. Knockdown of PTEN in PTEN positive cells leads to nucleolar morphologic changes and an increase in the proportion of cells with a greater number of nucleoli. In addition, knockdown of PTEN in PTEN positive cells increased ribosome biogenesis. These findings expand current understanding of function and relevance of nuclear localized PTEN and provide a foundation for the development of novel therapies targeting PTEN.     

Structural studies of a human hybrid immunoglobulin

The structure of an hypothesized hybrid or “Lepore-type” IgG contained in the serum of donor 2904 was investigated. Previous immunological studies suggested that this serum contained IgG with aγ3-γ1 hybrid heavy chain consisting of aγ3 Fd portion and aγ1 Fc. Structural studies have now shown that the carboxyl terminal end of the 2904 hybrid IgG, including much of the Fc fragment, isγ1 in character. However, the fingerprints of the Fc tryptic peptides at both pH 3.6 and pH 6.4 included a peptide, possibly from the hinge region, in peptide maps of Fc from aγG3 Gm (5) myeloma protein, but absent from maps of Fc peptides fromγG1 proteins. Gel filtration of the CNBr fragments of Fc from 2904 suggested that the hinge region isγ3-like. Papain cleavage experiments indicated an elevated level of resistant IgG, which agrees with immunological findings of an increasedγG2 subclass level. Our data confirm previous reports that theγ chain C-terminal octadecapeptides fromγG3 proteins have a subclass specific residue of arginine and indicate that within this subclass there is an allotypic variation related to the Gm type of the protein.     

Interrelationships between Gramineae lectins

Two Gramineae species, oat and maize, are compared with wheat and barley to see if they contain lectins which are structurally and functionally similar to the Hordeae lectins. Four distinct criteria were examined: localisation of lectin activity in the seed, ability to agglutinate a defined type of erythrocyte in a reaction reversed by monomers or oligomers of n-acetyl-d-glucosamine, ability to bind to the affinity matrix p-aminobenzyl-1-thio-2-acetamido-2-deoxy-β-d-glucopyranoside-substituted Sepharose, and cross-reactivity with monospecific antisera raised to wheat-germ agglutinin. Results indicate that the very close relationship found between the lectins of wheat, barley and rye cannot be extended to those species of Gramineae outside the tribe Hordeae.     

Changes in cell surface anionogenic groups during differentiation ofHerpetomonas samuelpessoai mediated by dimethylsulfoxide

The surface anionic groups of untreated or dimethyl sulfoxide (DMSO)-treatedHerpetomonas samuelpessoai cells were analyzed by cell electrophoresis, ultrastructural cytochemistry, and identification of sialic acids using thin-layer chromatography. Differentiation ofH. samuelpessoai induced by DMSO treatment caused a significant increase in the net negative surface charge. In flagellates exposed to DMSO, more cationized ferritin, colloidal iron hydroxide, and sendai virus particles bound to the cell surface. Treatment of both untreated and DMSO-treated flagellates with neuraminidase decreased markedly the EPM of cells to the cathodic pole. These findings suggest that sialic acid residues are the major anionogenic groups exposed on the surface ofH. samuelpessoai. Thin-layer chromatography showed thatN-acetyl andN,O-diacylneuraminic acids, in equal proportions, were present inH. samuelpessoai. However,N-acetylneuraminic acid predominates in DMSO-treated cells.     

Polymorphism of DNA Repair Gene xrcc1 and Lung Cancer Risk

The current large-scale meta-analysis was performed to reach a reliable conclusion on the association between X-ray repair cross-complementing 1 (xrcc1) rs1799782 and the development of lung cancer. Studies that investigated the association between rs1799782 and lung cancer risk were identified by searching PubMed. We calculated odds ratio (OR) with corresponding 95 % confidence interval (CI) for Trp/Trp vs Arg/Arg, Trp/Trp + Arg/Trp vs Arg/Arg, and Trp/Trp vs Arg/Trp + Arg/Arg contrast models. Combining all 25 studies, we yielded three summary ORs: 1.07 (95 % CI 0.92–1.23) for Trp/Trp vs Arg/Arg, 0.93 (95 % CI 0.87–1.00) for Trp/Trp + Arg/Trp vs Arg/Arg, and 1.08 (95 % CI 0.94–1.25) for Trp/Trp vs Arg/Trp + Arg/Arg, suggesting rs1799782 was not associated with overall risk of lung cancer. Strikingly, a significantly deceased risk was found among Caucasian populations (Trp/Trp + Arg/Trp vs Arg/Arg, OR = 0.86, 95 % CI 0.76–0.97). This study confirms that xrcc1 rs1799782 may lower the risk of lung cancer among Caucasians.     

Purification and characterization of aβ-galactosidase fromTreponema phagedenis (Reiter strain)

A β-galactosidase was highly purified from a cellular extract ofTreponema phagedenis (Reiter strain) by ammonium sulfate precipitation and successive chromatography on Sepharose 6B and DEAE-Sephadex. The purified enzyme was homogeneous in SDS-polyacrylamide gel electrophoresis. The molecular weight estimated was 580,000. The optimal pH, ionic strength, and temperature were 6.5, 0.1, and 50°C, respecitvely. The enzyme was stable only at around pH 6.5 and at temperatures lower than 35°C. The enzyme was irreversibly inhibited byp-chloromercuribenzoate and divalent cations such as Hg²⁺, Cu²⁺, Zn²⁺, Cd²⁺, and Pb²⁺. The Km values forp-nitrophenyl-β-D-galactopyranoside,o-nitrophenyl-β-D-galactopyranoside, and lactose were 0.29, 0.36, and 5.4 mM, respectively.     

The amino-terminal sequence ofStreptomyces griseus carboxypeptidase

The amino-terminal sequence of carboxypeptidase fromStreptomyces griseus was determined using a new protocol for automatic Edman degradation that reduced background noise. The sequence of the first 48 residues is: Asp-Phe-Pro-Pro-Ala-Asp-Ser-Arg-Tyr-His-Asn-Tyr-Ala-Glu-Met-Asn-Ala-Ala-Ile-Asp-Ala-Arg-Ile-Ala-Ala-Asn-Pro-Ser-Ile-Met-Ser-Lys-Arg-Val-Ile-Gly-Lys-Thr-Tyr-Gln-Gly-(Arg)-Asp-Val-Ile-Ala-Val-Lys, which is homologous to that of other zinc-containing carboxypeptidase from vertebrate and invertebrate sources.     

tRNA synthesis in germinating pollen

The synthesis of tRNA was demonstrated in pollen ofNicotiana tabacum L. according to the incorporation of labeled uracil, adenosine and guanosine during 4 h of germination. tRNA was extracted from postribosomal supernatant and purified by polyacrylamide gel electrophoresis. The incorporation of guanosine was about 1.68 times higher than that of adenosine. This finding indicates that the whole tRNA chain is formed in pollen tubes.