Recognition, targeting, and hydrolysis of the λ O replication protein by the ClpP/ClpX protease

Malgorzata Gonciarz-Swiatek, Alicja Wawrzynow, Soo Jong Um, Brian A Learn, Roger McMacken, William L. Kelley, Costa Georgopoulos, Olaf Sliekers, Maciej Zylicz

Research output: Contribution to journalArticle

Abstract

It has previously been established that sequences at the C termini of polypeptide substrates are critical for efficient hydrolysis by the ClpP/ClpX ATP-dependent protease. We report for the bacteriophage λ O replication protein, however, that N-terminal sequences play the most critical role in facilitating proteolysis by ClpP/ClpX. The N-terminal portion of λ O is degraded at a rate comparable with that of wild type O protein, whereas the C-terminal domain of O is hydrolyzed at least 10-fold more slowly. Consistent with these results, deletion of the first 18 amino acids of λ O blocks degradation of the N-terminal domain, whereas proteolysis of the O C-terminal domain is only slightly diminished as a result of deletion of the C-terminal 15 amino acids. We demonstrate that ClpX retains its capacity to bind to the N-terminal domain following removal of the first 18 amino acids of O. However, ClpX cannot efficiently promote the ATP-dependent binding of this truncated O polypeptide to ClpP, the catalytic subunit of the ClpP/ClpX protease. Based on our results with λ O protein, we suggest that two distinct structural elements may be required in substrate polypeptides to enable efficient hydrolysis by the ClpP/ClpX protease: (i) a ClpX-binding site, which may be located remotely from substrate termini, and (ii) a proper N- or C-terminal sequence, whose exposure on the substrate surface may be induced by the binding of ClpX.

Original languageEnglish (US)
Pages (from-to)13999-14005
Number of pages7
JournalJournal of Biological Chemistry
Volume274
Issue number20
DOIs
StatePublished - May 14 1999

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Hydrolysis
Peptide Hydrolases
Proteolysis
Amino Acids
Substrates
ATP-Dependent Proteases
Peptides
Proteins
Protein C
Bacteriophages
Catalytic Domain
Adenosine Triphosphate
Binding Sites
Degradation
polypeptide C

ASJC Scopus subject areas

  • Biochemistry

Cite this

Recognition, targeting, and hydrolysis of the λ O replication protein by the ClpP/ClpX protease. / Gonciarz-Swiatek, Malgorzata; Wawrzynow, Alicja; Um, Soo Jong; Learn, Brian A; McMacken, Roger; Kelley, William L.; Georgopoulos, Costa; Sliekers, Olaf; Zylicz, Maciej.

In: Journal of Biological Chemistry, Vol. 274, No. 20, 14.05.1999, p. 13999-14005.

Research output: Contribution to journalArticle

Gonciarz-Swiatek, M, Wawrzynow, A, Um, SJ, Learn, BA, McMacken, R, Kelley, WL, Georgopoulos, C, Sliekers, O & Zylicz, M 1999, 'Recognition, targeting, and hydrolysis of the λ O replication protein by the ClpP/ClpX protease', Journal of Biological Chemistry, vol. 274, no. 20, pp. 13999-14005. https://doi.org/10.1074/jbc.274.20.13999
Gonciarz-Swiatek, Malgorzata ; Wawrzynow, Alicja ; Um, Soo Jong ; Learn, Brian A ; McMacken, Roger ; Kelley, William L. ; Georgopoulos, Costa ; Sliekers, Olaf ; Zylicz, Maciej. / Recognition, targeting, and hydrolysis of the λ O replication protein by the ClpP/ClpX protease. In: Journal of Biological Chemistry. 1999 ; Vol. 274, No. 20. pp. 13999-14005.
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AU - McMacken, Roger

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