H-2M3 encodes the MHC class I molecule presenting the maternally transmitted antigen of the mouse

Mta, the maternally transmitted antigen of mice, is a hydrophobic, N-formylated mitochondrial peptide, MTF, presented on the cell surface to cytotoxic T lymphocytes by a novel major histocompatibility complex class I molecule, encoded by H-2M3. We have cloned and sequenced two alleles of M3, which differ in their ability to present MTF despite greater than 99% identity in the coding regions. M3 is as divergent from classical, antigen-presenting H-2 molecules as from other class I genes of the Hmt and the Qa/Tla regions. Amino acids critical for folding of class I molecules are conserved in M3. Noncharged amino acids lining the peptide-binding groove and phenylalanine 171 may explain the unique interaction with MTF, and leucine 95 appears critical for immunological activity.     

Purification and Characterization of d-Aminoacylase from Alcaligenes faecalis DA1

A d-aminoacylase from Alcaligenes faecalis DA1 has been purified to homogeneity by a simple purification procedure with two columns, Fractogel DEAE-650 and HW-50. The specific activity of the purified enzyme was found to be 580 U/mg of protein with N-acetyl-dl-methionine as the reaction substrate. The apparent molecular weight and isoelectric point of this enzyme were determined to be 55,000 and 5.4, respectively.     

Kinetic resolution of N-protected amino acid esters in organic solvents catalyzed by a stable industrial alkaline protease

Summary An industrial alkaline protease Alcalase has been found to be very stable in organic solvents and usable as a catalyst for resolution of N-protected amino acids, in both aqueous solution and organic solvent with high yield and optical purity. Only the L-amino acid ester has been hydrolysed.Abbreviation Cbz- carbobenzyloxy- - OMe methyl ester - Hop homophenylalanine - Nol norleucine - Aba -amino butyric acid - Nov norvaline - Fug furylglycine     

Expression and stability of amplified genes encoding 5-enolpyruvylshikimate-3-phosphate synthase in glyphosate-tolerant tobacco cells

Two distinct cDNAs for 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) were obtained from a glyphosate-tolerant tobacco cell line. The cDNAs were 89% identical and the predicted sequences of the mature proteins were greater than 83% identical with EPSPS proteins from other plants. Tobacco EPSPS proteins were more similar to those from tomato and petunia than Arabidopsis. One cDNA clone, EPSPS-1, represented a gene that was amplified in glyphosate-tolerant cells, while the gene for EPSPS-2 was unaltered in these cells. Consequently, EPSPS-1 mRNA was more abundant in tolerant than unselected cells, whereas EPSPS-2 mRNA was at relatively constant levels in these cell lines. Exposure of unselected cells and tobacco leaves to glyphosate produced a transient increase in EPSPS mRNA. However, glyphosate-tolerant cells containing amplified copies of EPSPS genes did not show a similar response following exposure to glyphosate. A significant proportion of the EPSPS gene amplification was maintained when tolerant cells were grown in the absence of glyphosate for eight months. Plants regenerated from these cells also contained amplified EPSPS genes.     

Artificial Induction and Evaluation of a Mild Isolate of Tomato Spotted Wilt Virus

A severe isolate (BL) of tomato spotted wilt virus (TSWV) that originated from Hawaii was treated with nitrous acid in an effort to obtain mild mutants. The standardized procedure used in mutation experiments was: extracting infected Gomphrena globosa L. leaf tissue in 0.01 M Na₂SO₃, 0.125 M sodium acetate and 0.4 M sodium nitrite at pH 5.5 and incubating the extract for 20 min at room temperature. The extract was inoculated to tobacco (Nicotiana tabacum L. cv. Havana 423) and local lesions were subsequently transferred to lettuce (Lactuca sativa L. cv. Minetto). One isolate (R27G) that incited mild symptoms in lettuce was obtained out of 868 local-lesion-transfers. Under greenhouse conditions, the isolate induced mild symptoms on tomato (Lycopersicon esculentum Mill.) but was severe on peppers (Capsicum annuum L.). The effect of the R27G isolate on growth of potted tomatoes kept outdoors was variable. In one trial, only 15 % of the fruit had symptoms versus 67 % in another trial. R27G fully protected Datura stramonium L. plants that were challenge inoculated with the severe parent BL isolate. Less effective cross protection was observed against a severe isolate from Oklahoma.     

Characterization of a human cervical carcinoma-associated antigen by lectin binding and immuno-electron microscopy

Summary The specific binding and nature of the epitope recognized by monoclonal antibody (Mab) 1H10, which binds an antigen expressed on human cervical tumors, was characterized by enzyme digestion, lectin competition assay and immuno-electron microscopy. Membrane homogenates of CaSki cervical carcinoma cells were digested with various enzymes, then analysed by SDS-PAGE and immunoblotting. Cells grown on coverslips were treated with various enzymes and in situ binding of Mab 1H10 to cells was analysed by electron microscopy. The ability of lectin-conjugates to block Mab 1H10 binding to CaSki cells was also examined. Treatment of samples with sodium periodate abrogated antigen recognition by Mab 1H10. Neuraminidase and hyaluronidase digestion decreased but did not eliminate Mab 1H10 binding to cells in situ. Chondroitinase ABC digestion, in contrast, removed Mab 1H10 binding sites both in vitro and in situ. Trypsin and chymotrypsin digestion of cell membrane homogenates decreased the molecular weight of the Mab 1H10 antigen but did not decrease the binding intensity. Wheat germ agglutinin (WGA) strongly bound to CaSki cells and partially blocked Mab 1H10 binding, indicating that the antigen contains N-acetyl-galactosamine residues at or near the epitope recognized by Mab 1H10. Ricinus communis agglutinin (RCA) exhibited a similar binding pattern to WGA. However, concanavalin A bound only weakly to CaSki cells and was ineffective at blocking Mab 1H10 binding. The tumor-associated antigen recognized by Mab 1H10 is concluded to be a chondroitin sulphate glycoprotein or proteoglycan rather than a mucopolysaccharide or lipoprotein.     

The plasma membrane calcium pump: a multiregulated transporter

Activation of many cells, especially nonexcitable cells, results in a Ca(2+) transient that is influenced in part by the kinetics of active extrusion of Ca(2+) across the plasma membrane. The molecular cloning of the plasma membrane Ca(2+)-pump has helped to clarify the relationship between its structure and function. The Ca(2+)-pump is controlled by multiple regulators, including calmodulin, phospholipids and various kinases. Longer term control is achieved through regulation of its gene expression, and the presence of a number of Ca(2+)-pump isoforms that differ in their regulatory domains provides potential functional diversity. In this review, we focus on the mechanisms that regulate the function of the Ca(2+)-pump, and their physiological significance.     

克氏鱼(Clarkeosteus)的新发现

文中记述了采自云南盘溪地区一真节甲鱼类化石,根据材料特征认为属于克氏鱼属—新种,即一打得克氏鱼(Clarkeosteus yidadeinsis sp. nov.).