Identification of a gene required for maturation of an extracellular lactococcal serine proteinase
| Autor: | Wn Konings, Alfred J. Haandrikman, Aat M. Ledeboer, Harry Laan, Gerhardus Venema, Jan Kok, S Soemitro |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 1989 |
| Předmět: |
DNA
Bacterial viruses genetic processes Molecular Sequence Data Restriction Mapping information science Enzyme-Linked Immunosorbent Assay Biology medicine.disease_cause Microbiology Bacterial Proteins Proteinase 3 medicine Amino Acid Sequence Molecular Biology Escherichia coli Gene Peptide sequence Base Sequence Lactococcus lactis Serine Endopeptidases Nucleic acid sequence RNA Streptococcus biochemical phenomena metabolism and nutrition biology.organism_classification Milk Proteins Molecular biology Enzyme Activation Molecular Weight Open reading frame Biochemistry bacteria Protein Processing Post-Translational Research Article Plasmids |
| Zdroj: | Journal of Bacteriology, 171(5), 2789-2794. AMER SOC MICROBIOLOGY |
| ISSN: | 0021-9193 |
| Popis: | Directly upstream of the Lactococcus lactis subsp. cremoris Wg2 proteinase gene is an oppositely directed open reading frame (ORF1). The complete nucleotide sequence of ORF1, encoding a 33-kilodalton protein, was determined. A protein of approximately 32 kilodaltons was synthesized when ORF1 was expressed in Escherichia coli by using a T7 RNA polymerase-specific promoter. L. lactis subsp. lactis MG1363 transformants carrying the proteinase gene but lacking ORF1 were phenotypically proteinase deficient, unlike transformants carrying both the proteinase gene and ORF1. Synthesis and secretion of proteinase antigen by L. lactis could be detected with proteinase-directed monoclonal antibodies regardless of whether ORF1 was present. The requirement of ORF1 for proteinase activation was reflected in a reduction in the molecular weight of the secreted proteinase. Furthermore, deletion of the 130 C-terminal amino acids of the Wg2 proteinase prevented attachment of the enzyme to lactococcal cells. |
| Databáze: | OpenAIRE |
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