title: Isolation and characterisation of a 13.8-kDa bacteriolytic enzyme from house dust mite extracts: homology with prokaryotic proteins suggests that the enzyme could be bacterially derived.
abstract: Bacteriolytic activity was detected in extracts of whole mite and spent growth medium (SGM) from the clinically important Dermatophagoides pteronyssinus and Dermatophagoides farinae mites and was most abundant in whole mite extract. Gram-positive organisms Micrococcus lysodeikticus, Bacillus megaterium and Listeria monocytogenes were preferentially lysed and the lytic activity was enhanced by thiols, destroyed by mite proteases, inhibited by HgCl2 and high concentrations of NaCl but was resistant to heat and acid treatment. Substrate SDS-PAGE analysis indicated the presence of several lytic enzymes, two of which were isolated from D. pteronyssinus spent growth medium extract by hydroxyapatite chromatography. The N-terminal amino acid sequence of one of them was then used in PCR-based cloning studies. The complete amino acid sequence of this protein was determined and cDNA found to encode a 130-amino acid residue mature protein with a 20-amino acid leader sequence. The deduced protein demonstrated sequence similarity with the C-terminal regions of a group of bacterial proteins belonging to the P60 superfamily. These data suggest that the enzyme is derived from bacteria within the mites rather than from mites per se.
authors.affiliation: Division of Inflammation and Infectious Diseases, Department of Microbiology, The University of Western Australia, 6009, Crawley, Australia.
authors.forename: Leslie T
authors.initials: LT
authors.lastname: Mathaba
authors.forename: Catherine H
authors.initials: CH
authors.lastname: Pope
authors.forename: Jason
authors.initials: J
authors.lastname: Lenzo
authors.forename: Maria
authors.initials: M
authors.lastname: Hartofillis
authors.forename: Helen
authors.initials: H
authors.lastname: Peake
authors.forename: Robert L
authors.initials: RL
authors.lastname: Moritz
authors.forename: Richard J
authors.initials: RJ
authors.lastname: Simpson
authors.forename: Andreas
authors.initials: A
authors.lastname: Bubert
authors.forename: Philip J
authors.initials: PJ
authors.lastname: Thompson
authors.forename: Geoffrey A
authors.initials: GA
authors.lastname: Stewart
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D001433:Bacteriolysis; D001483:Base Sequence; D017077:Culture Media, Conditioned / Q000737:chemistry / Q000378:metabolism; D018076:DNA, Complementary; D004391:Dust; D004591:Electrophoresis, Polyacrylamide Gel; D004798:Enzymes / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D006094:Gram-Positive Bacteria / Q000201:enzymology / Q000502:physiology*; D006798:Housing; D008925:Mites / Q000201:enzymology* / Q000382:microbiology; D008969:Molecular Sequence Data
chemical_list: D001426:Bacterial Proteins; D017077:Culture Media, Conditioned; D018076:DNA, Complementary; D004391:Dust; D004798:Enzymes