Active-Site-Directed Chemical Tools for Profiling Mitochondrial Lon Protease

Fishovitz, Jennifer; Li, Min; Frase, Hilary; Hudak, Jason E.; Craig, Sandra; Ko, Kristin; Berdis, Anthony J.; Suzuki, Carolyn K.; Lee, Irene

doi:10.1021/cb100408w

Cited by 14 publications

(30 citation statements)

References 33 publications

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“…Mammalian homolog of Lon, together with mammalian homolog of another ATP-dependent bacterial serine protease, ClpXP, is involved in the protein quality control in the mitochondrial matrix. Peptidyl boronates are capable of inhibiting mammalian Lon and ClpXP proteases (Fishovitz et al, 2011), although inhibition of these proteases by bortezomib or two boronates in clinical trials has not been reported.…”

Section: Major Structural Classes Of Inhibitors Of Proteolytic Sites mentioning

confidence: 99%

Proteasome Inhibitors: An Expanding Army Attacking a Unique Target

Kisselev

Linden

Overkleeft

2012

Chemistry & Biology

480

482

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Proteasomes are large, multisubunit proteolytic complexes presenting multiple targets for therapeutic intervention. The 26S proteasome consists of a 20S proteolytic core and one or two 19S regulatory particles. The 20S core contains three types of active sites. Many structurally diverse inhibitors of these active sites, both natural product and synthetic, have been discovered in the last two decades. One, bortezomib, is used clinically for treatment of multiple myeloma, mantle cell lymphoma, and acute allograft rejection. Five more recently developed proteasome inhibitors are in trials for treatment of myeloma and other cancers. Proteasome inhibitors also have activity in animal models of autoimmune and inflammatory diseases, reperfusion injury, promote bone and hair growth, and can potentially be used as anti-infectives. In addition, inhibitors of ATPases and deubiquitinases of 19S regulatory particles have been discovered in the last decade.

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Section: Major Structural Classes Of Inhibitors Of Proteolytic Sites mentioning

confidence: 99%

Proteasome Inhibitors: An Expanding Army Attacking a Unique Target

Kisselev

Linden

Overkleeft

2012

Chemistry & Biology

480

482

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“…Upon cleavage of FRETN 89-98, the donor and acceptor separate from one another leading to increased fluorescence. Human mitochondrial Lon also degrades FRETN 89-98 and has been used to monitor its proteolytic activity [52]. Although FRETN 89-98 is also degraded by the 20S proteasome, this activity is not stimulated by ATP and thus can be distinguished from Lon [52].…”

Section: Enzymology Of Mitochondrial Lonmentioning

confidence: 99%

“…Human mitochondrial Lon also degrades FRETN 89-98 and has been used to monitor its proteolytic activity [52]. Although FRETN 89-98 is also degraded by the 20S proteasome, this activity is not stimulated by ATP and thus can be distinguished from Lon [52]. Additional peptide or chemical reporters are required to distinguish specific mitochondrial and cytosolic ATP-dependent protease activity in cellular extracts.…”

Section: Enzymology Of Mitochondrial Lonmentioning

confidence: 99%

“…However, insights into the functional mechanics of mitochondrial Lon can be drawn from the bacterial system as human Lon is able to degrade some bacterial Lon substrates in vitro [52], and yeast Lon/Pim1p can be functionally replaced by E. coli Lon [55]. Comparing the degradation profiles of several endogenous or heterologous substrates of E. coli Lon led to the postulate that solvent exposed hydrophobic peptide patches in an unfolded protein function as recognition tags for the protease [56–59].…”

Section: Enzymology Of Mitochondrial Lonmentioning

confidence: 99%

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Multitasking in the mitochondrion by the ATP-dependent Lon protease

Venkatesh

Lee

Singh

et al. 2012

Biochimica et Biophysica Acta (BBA) - Molecular Cell Research

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145

138

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The AAA+ Lon protease is a soluble single-ringed homo-oligomer, which represents the most streamlined operational unit mediating ATP-dependent proteolysis. Despite its simplicity, the architecture of Lon proteases exhibits a species-specific diversity. Homology modeling provides insights into the structural features that distinguish bacterial and human Lon proteases as hexameric complexes from yeast Lon, which is uniquely heptameric. The best-understood functions of mitochondrial Lon are linked to maintaining proteostasis under normal metabolic conditions, and preventing proteotoxicity during environmental and cellular stress. An intriguing property of human Lon is its specific binding to G-quadruplex DNA, and its association with the mitochondrial genome in cultured cells. A fraction of Lon preferentially binds to the control region of mitochondrial DNA where transcription and replication are initiated. Here, we present an overview of the diverse functions of mitochondrial Lon, as well as speculative perspectives on its role in protein and mtDNA quality control.

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“…Other protective factors in the presence of MPP+ include elevation of lon peptidase 1 which would strengthen mitochondria by enhancing degradation of damaged proteins by ROS or other types of stressors in the matrix. (Bayot et al, 2008, Fishovitz et al, 2011, Pinti et al, 2011, Wagatsuma et al, 2011) In summary, the collective general changes in the transcriptome which impacted the mitochondria are associated with either 1) primary loss of OXPHOS genes or 2) rise in systems that attenuate mitochondrial induced apoptosis.…”

Section: 0 Discussionmentioning

confidence: 99%

Whole genome expression profile in neuroblastoma cells exposed to 1-methyl-4-phenylpyridine

Mazzio

Soliman

2012

NeuroToxicology

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Mitochondrial dysfunction and subsequent energy failure is a contributing factor to degeneration of the substantia nigra pars compacta associated with Parkinson’s disease (PD). In this study, we investigate molecular events trigger by 1-methyl-4-phenylpyridine (MPP+) using whole genome-expression microarray, western blot and HPLC quantification of metabolites. The data show that MPP+ (500μM) evokes obstruction of mitochondrial respiration/oxidative phosphorylation (OXPHOS) in mouse neuroblastoma Neuro-2a cells, juxtapose to accelerated glucose consumption and production of lactic acid. While additional glucose concentrations restored viability at MPP+ (500μM), loss of OXPHOS was sustained suggesting that compensatory anaerobic metabolic systems were fulfilling required energy needs. Under these conditions, MPP+ initiated significant changes to the transcription of 799 genes (596 up-regulated and 203 down-regulated) of which 612 David IDS were applied and 136 functional annotation clusters were identified. Prominent changes were as follows; MPP+ initiated loss of mRNA for mitochondrial encoded NADH dehydrogenase 4, 5 genes, cytochrome b and NADH dehydrogenase (ubiquinone) flavoprotein 3, concomitant to rise in a mitochondrial fission gene; ganglioside-induced differentiation-associated-protein 1 (GDAP1). These negative changes to OXPHOS components were accompanied by a number of protective forces to the mitochondria including elevated ratio of mitochondrial anti/pro-apoptotic processes including a loss of apoptotic Bcl-2/adenovirus E1B 19-kDa-interacting protein (BNIP3) and family with sequence similarity 162, member A (FAM162a) and rise of heat shock protein 1 and Lon peptidase 1. There were no changes indicative of free radical damage (e.g. SOD, GSH-Px), rather MPP+ initiated a large number of significant G protein signaling components (which trigger catabolic processes) and anaerobic metabolic systems involving carboxylic acid/ transamination reactions (e.g. glutamate oxaloacetate transaminase 1 (GOT1), glutamic pyruvate-alanine transaminase 2 (GPT2), cystathionase and redox proteins such as cytochrome b5 reductase 1 and ferredoxin reductase. Counter-intuitively, the data show reduction of mRNA in glycolytic processes [DAVID enrichment score 9.96 p value 1.90E-19], some corroborated by western blot, bringing in to question the sources of lactate observed in the presence of MPP+. Examining this aspect, the data show that diverse carboxylic acids (succinate, oxaloacetate and a-ketoglutarate) are capable of contributing to the lactate pool in addition to phosph(enolpyruvate) or pyruvate in the absence of glucose by this cell line. In conclusion, these findings show that MPP+ negatively affects the transcriptome involved with complex I, but evoked elevation in G protein signaling and anaerobic metabolic systems involved with nitrogen/carboxylic acid metabolism and redox reactions. Anaerobic survival systems appear to be far more complex than previously believed, and future research will be required to elucidate th...

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Active-Site-Directed Chemical Tools for Profiling Mitochondrial Lon Protease

Cited by 14 publications

References 33 publications

Proteasome Inhibitors: An Expanding Army Attacking a Unique Target

Proteasome Inhibitors: An Expanding Army Attacking a Unique Target

Multitasking in the mitochondrion by the ATP-dependent Lon protease

Whole genome expression profile in neuroblastoma cells exposed to 1-methyl-4-phenylpyridine

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