Characterization of the HslU chaperone affinity for HslV protease

M. Kamran Azim, Walter Goehring, Hyun Kyu Song, Ravishankar Ramachandran, Matthias Bochtler, Peter Goettig

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

The HslVU complex is a bacterial two-component ATP-dependent protease, consisting of HslU chaperone and HslV peptidase. Investigation of protein-protein interactions using SPR in Escherichia coli HslVU and the protein substrates demonstrates that HslU and HslV have moderate affinity (Kd = 1 μM) for each other. However, the affinity of HslU for HslV fivefold increased (Kd ∼ 0.2 μM) after binding with the MBP-SulA protein indicating the formation of a "ternary complex" of HslV-HslU-MBP-SulA. The molecular interaction studies also revealed that HslU strongly binds to MBP-SulA with 10-9 M affinity but does not associate with nonstructured casein. Conversely, HslV does not interact with the MBP-SulA whereas it strongly binds with casein (Kd = 0.2 μM) requiring an intact active site of HslV. These findings provide evidence for "substrate-induced" stable HslVU complex formation. Presumably, the binding of HslU to MBP-SulA stimulates a conformational change in HslU to a high-affinity form for HslV.

Original languageEnglish
Pages (from-to)1357-1362
Number of pages6
JournalProtein Science
Volume14
Issue number5
DOIs
Publication statusPublished - 2005 May 1

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Peptide Hydrolases
Caseins
ATP-Dependent Proteases
Proteins
Escherichia coli Proteins
Molecular interactions
Catalytic Domain
Substrates
Escherichia coli

ASJC Scopus subject areas

  • Biochemistry

Cite this

Azim, M. K., Goehring, W., Song, H. K., Ramachandran, R., Bochtler, M., & Goettig, P. (2005). Characterization of the HslU chaperone affinity for HslV protease. Protein Science, 14(5), 1357-1362. https://doi.org/10.1110/ps.04970405

Characterization of the HslU chaperone affinity for HslV protease. / Azim, M. Kamran; Goehring, Walter; Song, Hyun Kyu; Ramachandran, Ravishankar; Bochtler, Matthias; Goettig, Peter.

In: Protein Science, Vol. 14, No. 5, 01.05.2005, p. 1357-1362.

Research output: Contribution to journalArticle

Azim, MK, Goehring, W, Song, HK, Ramachandran, R, Bochtler, M & Goettig, P 2005, 'Characterization of the HslU chaperone affinity for HslV protease', Protein Science, vol. 14, no. 5, pp. 1357-1362. https://doi.org/10.1110/ps.04970405
Azim MK, Goehring W, Song HK, Ramachandran R, Bochtler M, Goettig P. Characterization of the HslU chaperone affinity for HslV protease. Protein Science. 2005 May 1;14(5):1357-1362. https://doi.org/10.1110/ps.04970405
Azim, M. Kamran ; Goehring, Walter ; Song, Hyun Kyu ; Ramachandran, Ravishankar ; Bochtler, Matthias ; Goettig, Peter. / Characterization of the HslU chaperone affinity for HslV protease. In: Protein Science. 2005 ; Vol. 14, No. 5. pp. 1357-1362.
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