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.
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.
Mathaba
Leslie T
LT
Division of Inflammation and Infectious Diseases, Department of Microbiology, The University of Western Australia, 6009, Crawley, Australia.
Pope
Catherine H
CH
Lenzo
Jason
J
Hartofillis
Maria
M
Peake
Helen
H
Moritz
Robert L
RL
Simpson
Richard J
RJ
Bubert
Andreas
A
Thompson
Philip J
PJ
Stewart
Geoffrey A
GA
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
D001426:Bacterial Proteins; D017077:Culture Media, Conditioned; D018076:DNA, Complementary; D004391:Dust; D004798:Enzymes