Dataset Information

Interactome analysis by shotgun proteomics identifies a novel substrate for Mycolicibacterium smegmatis unfoldase ClpC1

ABSTRACT: Tuberculosis remains a leading cause of worldwide infectious mortality and one of the top ten leading causes of death overall. While advances in public health have contributed to a reduction in tuberculosis cases, the prevalence of multidrug-resistant Mycobacterium tuberculosis (Mtb) (MDR-TB) infections has created an urgent need to exploit novel drug targets. One such target is the ClpC1P1P2 protease, which degrades folded cytosolic proteins through the cooperation of the ATP-dependent unfoldase ClpC1 and the ClpP1P2 peptidase. Both protease components are strictly essential for Mtb viability and are validated therapeutic targets. However, efforts to develop anti-Mtb compounds are constrained by a limited understanding of Clp protease function and essentiality. Thus, it is crucial to identify physiological substrates and pathways regulated by this protease. In this study, we identify cellular proteins that interact with the ClpC1 unfoldase in Mycolicibacterium smegmatis (Msm), a nonpathogenic surrogate for Mtb. Using a FLAG-tagged ClpC1 variant with mutations within the Walker B motifs, candidate ClpC1 interaction partners were captured by co-immunoprecipitation and identified by mass spectrometry-based proteomics (LC-MS/MS). Notably, our work reveals a novel proteolytic substrate, 5-oxoprolinase, which is recognized by ClpC1P1P2 via an N-terminal degradation sequence.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Mycobacterium Smegmatis (strain Atcc 700084 / Mc(2)155)

SUBMITTER: Emmanuel Ogbonna

LAB HEAD: Karl Robert Schmitz

PROVIDER: PXD030385 | Pride | 2022-12-12

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	Schmitz_Emmanuel_C1D_15_1_191103060850.mgf	Mgf
	Schmitz_Emmanuel_C1D_15_1_191103060850.raw	Raw
	Schmitz_Emmanuel_C1D_15_2.mgf	Mgf
	Schmitz_Emmanuel_C1D_15_2.raw	Raw
	Schmitz_Emmanuel_C1D_15_2_191103094915.mgf	Mgf

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Publications

Interactome Analysis Identifies MSMEI_3879 as a Substrate of Mycolicibacterium smegmatis ClpC1.

Ogbonna Emmanuel C EC Anderson Henry R HR Beardslee Patrick C PC Bheemreddy Priyanka P Schmitz Karl R KR

Microbiology spectrum 20230621 4

The prevalence of drug-resistant Mycobacterium tuberculosis infections has prompted extensive efforts to exploit new drug targets in this globally important pathogen. ClpC1, the unfoldase component of the essential ClpC1P1P2 protease, has emerged as one particularly promising antibacterial target. However, efforts to identify and characterize compounds that impinge on ClpC1 activity are constrained by our limited knowledge of Clp protease function and regulation. To expand our understanding of C ...[more]

PMID: 37341639

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