Recombinant Deg/HtrA proteases from Synechocystis sp. PCC 6803 differ in substrate specificity, biochemical characteristics and mechanism
Recombinant Deg/HtrA proteases from Synechocystis sp. PCC 6803 differ in substrate specificity, biochemical characteristics and mechanism
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2011
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Huesgen, Pitter F.
Miranda, Helder
Lam, XuanTam
Funk, Christiane
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Biochemical Journal ; 435 (2011), 3. - pp. 733-742. - ISSN 0264-6021. - eISSN 1470-8728
Abstract
Cyanobacteria require efficient protein-quality-control mechanisms to survive under dynamic, often stressful, environmental conditions. It was reported that three serine proteases, HtrA (high temperature requirement A), HhoA (HtrA homologue A) and HhoB (HtrA homologue B), are important for survival of Synechocystis sp. PCC 6803 under high light and temperature stresses and might have redundant physiological functions. In the present paper, we show that all three proteases can degrade unfolded model substrates, but differ with respect to cleavage sites, temperature and pH optima. For recombinant HhoA, and to a lesser extent for HtrA, we observed an interesting shift in the pH optimum from slightly acidic to alkaline in the presence of Mg2+ and Ca2+ ions. All three proteases formed different homo-oligomeric complexes with and without substrate, implying mechanistic differences in comparison with each other and with the well-studied Escherichia coli orthologues DegP (degradation of periplasmic proteins P) and DegS. Deletion of the PDZ domain decreased, but did not abolish, the proteolytic activity of all three proteases, and prevented substrate-induced formation of complexes higher than trimers by HtrA and HhoA. In summary, biochemical characterization of HtrA, HhoA and HhoB lays the foundation for a better understanding of their overlapping, but not completely redundant, stress-resistance functions in Synechocystis sp. PCC 6803.
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570 Biosciences, Biology
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Biochemical characterization,oligomerization,PDZ domain,serine protease,substrate specificity,Synechocystis sp. PCC 6803
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HUESGEN, Pitter F., Helder MIRANDA, XuanTam LAM, Manuela PERTHOLD, Holger SCHUHMANN, Iwona ADAMSKA, Christiane FUNK, 2011. Recombinant Deg/HtrA proteases from Synechocystis sp. PCC 6803 differ in substrate specificity, biochemical characteristics and mechanism. In: Biochemical Journal. 435(3), pp. 733-742. ISSN 0264-6021. eISSN 1470-8728. Available under: doi: 10.1042/BJ20102131BibTex
@article{Huesgen2011-05-01Recom-19505, year={2011}, doi={10.1042/BJ20102131}, title={Recombinant Deg/HtrA proteases from Synechocystis sp. PCC 6803 differ in substrate specificity, biochemical characteristics and mechanism}, number={3}, volume={435}, issn={0264-6021}, journal={Biochemical Journal}, pages={733--742}, author={Huesgen, Pitter F. and Miranda, Helder and Lam, XuanTam and Perthold, Manuela and Schuhmann, Holger and Adamska, Iwona and Funk, Christiane} }
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