Molecular basis of interactions between mitochondrial proteins and hydroxyapatite in the presence of Triton X-100, as revealed by proteomic and recombinant techniques

Yamamoto, Toshiharu; Tamaki, Haruna; Katsuda, Chie; Nakatani, Kiwami; Terauchi, Satsuki; Terada, Hiroshi; Shinohara, Yasuo

doi:10.1016/j.chroma.2013.05.079

Cited by 9 publications

(14 citation statements)

References 43 publications

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“…To localize their binding site in AAC, SMPs (2.0 mg of proteins/mL) were incubated with 10 μM AMM-120 for 1 h, solubilized with 5% (w/v) Triton X-100, and passed through a hydroxyapatite column, followed by conjugation with a fluorescent TAMRA-N 3 tag via Cu + -catalyzed click chemistry. Hydroxyapatite chromatography provided AAC as an almost single band on a SDS gel (the control lane in Figure A), as reported previously. , …”

Section: Resultssupporting

confidence: 82%

“…Hydroxyapatite chromatography provided AAC as an almost single band on a SDS gel (the control lane in Figure 9A), as reported previously. 27,31 The purified AMM-120-bound AAC was digested by CNBr or lysylendopeptidase (Lys-C), and the digests were separated on a 16.5% Schagger-type tricine gel. CNBr cleavage provided a strong fluorescent band in the ∼20 kDa fragment (Figure 9A).…”

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confidence: 99%

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Epoxycyclohexenedione-Type Compounds Make Up a New Class of Inhibitors of the Bovine Mitochondrial ADP/ATP Carrier

et al. 2018

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Through the extensive screening of our chemical library, we found epoxycyclohexenedione (ECHD)-type compounds (AMM-59 and -120) as unique inhibitors of the bovine heart mitochondrial ADP/ATP carrier (AAC). This study investigated the mechanism of inhibition of AAC by ECHDs using submitochondrial particles (SMPs). Proteomic analyses of ECHD-bound AAC as well as biochemical characterization using different SH reagents showed that ECHDs inhibit the function of AAC by covalently binding primarily to Cys57 and secondarily to Cys160. Interestingly, AAC remarkably aggregated in SMPs upon being incubated with high concentrations of ECHDs for a long period of time. This aggregation was observed under both oxidative and reductive conditions of the sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of SMP proteins, indicating that aggregation is not caused by intermolecular S-S linkages. ECHDs are the first chemicals, to the best of our knowledge, to induce prominent structural alteration in AAC without forming intermolecular S-S linkages. When all solvent-accessible cysteines (Cys57, Cys160, and Cys257) were previously modified by N-ethylmaleimide, the aggregation of AAC was completely suppressed. In contrast, when Cys57 or Cys160 is selectively modified by a SH reagent, the covalent binding of ECHDs to a residual free residue of the two cysteines is sufficient to induce aggregation. The aggregation-inducing ability of another ECHD analogue (AMM-124), which has an alkyl chain that is shorter than those of AMM-59 and -120, was significantly less efficient than that of the two compounds. On the basis of these results, the mechanism underlying the aggregation of AAC induced by ECHDs is discussed.

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Section: Resultssupporting

confidence: 82%

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confidence: 99%

Epoxycyclohexenedione-Type Compounds Make Up a New Class of Inhibitors of the Bovine Mitochondrial ADP/ATP Carrier

et al. 2018

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“…Rat liver mitochondria were solubilized with Triton X-100 and chromatographed through a hydroxyapatite (HTP) column [ 34 ]. Since there is no established protocol for the purification of VDAC3 from tissues, the protein is enriched in the HTP eluate [ 35 ] that contains several electrophoretic bands in the range of 30-35 kDa [ 35 ]. The HTP eluate was either reduced or oxidized and run on a gel.…”

Section: Resultsmentioning

confidence: 99%

VDAC3 as a sensor of oxidative state of the intermembrane space of mitochondria: the putative role of cysteine residue modifications

et al. 2016

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“…The Arabidopsis thaliana genome encodes four Coq11 orthologs (At1g32220, At5g15910, At5g15480, and At5g10730), and the chloroplast-localized flavin reductase-related protein, At1g32220, is thought to be involved in plastoquinone biosynthesis (49,78). In COXPRESdb, COQ10A is coexpressed with SDR39U1, which, like Coq11, belongs to the SDR superfamily (75), and SDR39U1 was found to coelute with human COQ9 from a hydroxyapatite column in mitochondria solubilized with Triton X-100 (91). These functional inferences may supply a link between human COQ10 and Coq11-like proteins in CoQ production.…”

Section: Discussionmentioning

confidence: 99%

Human COQ10A and COQ10B are distinct lipid-binding START domain proteins required for coenzyme Q function

Tsui

Pham

Amer

et al. 2019

Journal of Lipid Research

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Coenzyme Q (ubiquinone or Q) functions as an essential redox‐active lipid in respiratory electron and proton transport in cellular energy metabolism, and it is an important lipid‐soluble antioxidant in cellular membranes. In Saccharomyces cerevisiae, proteins Coq3‐Coq9, as well as Coq11, assemble into a multi‐subunit protein complex called the CoQ‐synthome, which is required for Q biosynthesis. Coq10, a putative steroidogenic acute regulatory (StAR)‐related lipid transfer (StART) domain protein is not a member of the CoQ‐synthome, but is required for the proper assembly of the CoQ‐synthome, efficient de novo Q biosynthesis during early‐log phase, and the function of Q in respiration and as an antioxidant. Humans possess two isoforms, namely COQ10A and COQ10B. Based on the RNA‐seq data from NCBI, COQ10A is predominantly expressed in heart while COQ10B is present in all tissues, suggesting that COQ10B is probably serving a more general role. Previous studies have shown rescue of S. cerevisiae coq10Δ respiration deficient phenotype by expression of human COQ10A. Here we present new evidence showing rescue of S. cerevisiae coq10Δ by expression of either the human COQ10A or COQ10B homolog, as determined by restoration of respiratory growth on non‐fermentable carbon sources, de novo Q biosynthesis, as well as restoration of the CoQ‐synthome. Support or Funding Information This research was supported by NSF MCB‐1330803 and Ruth L. Kirschstein National Research Service Award GM007185. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

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Molecular basis of interactions between mitochondrial proteins and hydroxyapatite in the presence of Triton X-100, as revealed by proteomic and recombinant techniques

Cited by 9 publications

References 43 publications

Epoxycyclohexenedione-Type Compounds Make Up a New Class of Inhibitors of the Bovine Mitochondrial ADP/ATP Carrier

Epoxycyclohexenedione-Type Compounds Make Up a New Class of Inhibitors of the Bovine Mitochondrial ADP/ATP Carrier

VDAC3 as a sensor of oxidative state of the intermembrane space of mitochondria: the putative role of cysteine residue modifications

Human COQ10A and COQ10B are distinct lipid-binding START domain proteins required for coenzyme Q function

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