Dissecting capture and twisting of aureolysin and pseudolysin: functional amino acids of the Dispase autolysis-inducing protein.

Abstract

The Dispase autolysis-inducing protein (DAIP) from Streptomyces mobaraensis attracts M4 metalloproteases, which results in inhibition and autolysis of bacillolysin (BL) and thermolysin (TL). The present study shows that aureolysin (AL) from Staphylococcus aureus and pseudolysin (LasB) from Pseudomonas aeruginosa are likewise impaired by DAIP. Complete inhibition occurred when DAIP significantly exceeded the amount of the target protease. At low DAIP concentrations, AL and BL performed autolysis, while LasB and TL degradation required reductants or detergents that break intramolecular disulfide bonds or change the protein structure. Site directed mutagenesis of DAIP and removal of an exposed protein loop either influenced binding or inhibition of AL and TL but had no effect on LasB and BL. The Y170A and Δ239-248 variants had completely lost affinity for TL and AL. The exchange of Asn-275 also impaired the interaction of DAIP with AL. In contrast, DAIP Phe-297 substitution abolished inhibition and autolysis of both target proteases but still allowed complex formation. Our results give rise to the conclusion that other, yet unknown DAIP amino acids inactivate LasB and BL. Obviously, various bacteria in the same habitat caused Streptomyces mobaraensis to continuously optimize DAIP in inactivating the tackling metalloproteases.

• Authors: David Fiebig, Anita Anderl, Sahra Al Djaizani, Harald Kolmar, Hans-Lothar Fuchsbauer
• Journal: The Biochemical journal (Biochem J) Vol. 477 Issue 13 Pg. 2595-2606 (07 17 2020) ISSN: 1470-8728 [Electronic] England
• PMID: 32602533 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2020 The Author(s). Published by Portland Press Limited on behalf of the Biochemical Society.

Chemical References

• Amino Acids
• Bacterial Proteins
• Endopeptidases
• Metalloproteases
• Metalloendopeptidases
• aureolysin
• pseudolysin, Pseudomonas aeruginosa
• dispase

Topics

• Amino Acids (chemistry, metabolism)
• Autolysis (metabolism)
• Bacterial Proteins (chemistry, metabolism)
• Calorimetry
• Chromatography, Gel
• Circular Dichroism
• Endopeptidases (chemistry, metabolism)
• Metalloendopeptidases (chemistry, metabolism)
• Metalloproteases (chemistry, metabolism)
• Staphylococcus aureus (enzymology)