Apolipophorin III is a substrate for protease IV from Pseudomonas aeruginosa.
Our results demonstrated that Pseudomonas aeruginosa serine protease IV degraded apolipophorin III from the haemolymph of Galleria mellonella larvae. ApoLp-III protein was degraded in a stepwise manner. Four intermediate forms of 15, 13.3, 11.9 and 9.5 kDa were detected after 30 min digestion while only one of 5.6 kDa was released after 1-h incubation time. N-terminal amino acid sequence analysis of 5.6 kDa peptide revealed that it was released from apoLp-III after cleavage between lysine 70 and 71. ApoLp-III degradation by protease IV was inhibited by 1 mM TLCK but not 1 mM EDTA, additionally demonstrating that digestion was catalysed by a serine protease. Our data also indicated apoLp-III degradation in vivo during P. aeruginosa infection of G. mellonella larvae.
Andrejko
Mariola
M
Department of Invertebrate Immunology, Maria Curie-Skłodowska University, Akademicka 19, 20-033 Lublin, Poland.
Cytryńska
Małgorzata
M
Jakubowicz
Teresa
T
D000595:Amino Acid Sequence; D000818:Animals; D001053:Apolipoproteins / Q000378:metabolism*; D006458:Hemolymph / Q000378:metabolism; D006801:Humans; D007814:Larva / Q000382:microbiology; D008969:Molecular Sequence Data; D009036:Moths / Q000382:microbiology*; D010447:Peptide Hydrolases / Q000378:metabolism*; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D013379:Substrate Specificity
D001053:Apolipoproteins; C042470:apolipophorin III; D010447:Peptide Hydrolases; C030732:protease IV