Abstract |
An acidic endoprotease (MAEP) secreted during vegetative growth by Myxococcus xanthus DK101 was purified to homogeneity by a series of chromatographic procedures. The endoprotease cleaved the Phe-Met bond of kappa-casein under acidic conditions (pH 5.9). Its apparent molecular mass and its isoelectric point have been estimated to be 12 kDa and 4.5, respectively. From the N-terminal amino acid sequence, a set of two primers for polymerase chain reaction have been designed. Amplification of the corresponding DNA fragment (84 bp) generated a probe, then used to screen an expression DNA library of M. xanthus and to isolate a recombinant plasmid which contained a 2127-bp insert. The nucleotide sequence included an open reading frame (ORF) of 585 nucleotides, encoding 195 amino acids, that exhibited a high degree of similarity with the N-terminal amino acid sequence of the purified MAEP. The polypeptide sequence inferred from this ORF revealed that the mature enzyme should contain 131 amino acids arising from a 195-amino-acid precursor protein.
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Authors | N Lucas, C Mazaud-Aujard, L Bremaud, Y Cenatiempo, R Julien |
Journal | European journal of biochemistry
(Eur J Biochem)
Vol. 222
Issue 2
Pg. 247-54
(Jun 01 1994)
ISSN: 0014-2956 [Print] England |
PMID | 8020464
(Publication Type: Comparative Study, Journal Article)
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Chemical References |
- DNA Primers
- DNA, Bacterial
- Endopeptidases
- Myxococcus acidic endoprotease
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Topics |
- Amino Acid Sequence
- Base Sequence
- Chromatography, Gel
- Chromatography, High Pressure Liquid
- Chromatography, Ion Exchange
- Chromosomes, Bacterial
- Cloning, Molecular
- DNA Primers
- DNA, Bacterial
(chemistry, isolation & purification)
- Endopeptidases
(genetics, isolation & purification, metabolism)
- Genes, Bacterial
- Molecular Sequence Data
- Molecular Weight
- Myxococcus xanthus
(enzymology, genetics)
- Open Reading Frames
- Polymerase Chain Reaction
- Promoter Regions, Genetic
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