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Identification of dGTP-interacting mitochondrial proteins by affinity purification and LC-ESI-MS/MS

ABSTRACT: Imbalanced mitochondrial dNTP pools are known players in the pathogenesis of multiple human diseases. However, we have shown that dGTP is largely overrepresented over the other dNTPs in mitochondria of mouse tissues and human cultured cells. Here, we have performed affinity purification studies and identified NDUFA10, an accessory subunit of respiratory complex I, as the protein binding most of the dGTP contained in mitochondria. This interaction provides a possible link between oxidative metabolism and regulation of dNTP availability and thus with DNA maintenance.

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Liver

SUBMITTER: Yolanda Cámara

LAB HEAD: Ramon Martí

PROVIDER: PXD033900 | Pride | 2022-06-29

REPOSITORIES: Pride

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			Action	DRS
	Band_1.baf	Other
	Band_1.dat	Other
	Band_1.mgf	Mgf
	Band_2.baf	Other
	Band_2.dat	Other

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Publications

Most mitochondrial dGTP is tightly bound to respiratory complex I through the NDUFA10 subunit.

Molina-Granada David D González-Vioque Emiliano E Dibley Marris G MG Cabrera-Pérez Raquel R Vallbona-Garcia Antoni A Torres-Torronteras Javier J Sazanov Leonid A LA Ryan Michael T MT Cámara Yolanda Y Martí Ramon R

Communications biology 20220623 1

Imbalanced mitochondrial dNTP pools are known players in the pathogenesis of multiple human diseases. Here we show that, even under physiological conditions, dGTP is largely overrepresented among other dNTPs in mitochondria of mouse tissues and human cultured cells. In addition, a vast majority of mitochondrial dGTP is tightly bound to NDUFA10, an accessory subunit of complex I of the mitochondrial respiratory chain. NDUFA10 shares a deoxyribonucleoside kinase (dNK) domain with deoxyribonucleosi ...[more]

PMID: 35739187

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2024-09-09 | PXD051958 | Pride

Identification of novel KRT32-associated proteins in primary keratinocytes cells lysate by GST-KRT32 pulldown assays coupled with mass spectrometry.

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Identification of proteins associated with the human mitochondrial degradosome

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Profiling of human spermatozoa glycoproteins by LC/MS/MS

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Proteomics Reveals Extracellular Matrix Injury in the Glomeruli and Tubulointerstitium of Kidney Allografts with Early Antibody-Mediated Rejection

Project description:Antibody-mediated rejection (AMR) accounts for >50% of kidney allograft losses. AMR is caused by donor-specific antibodies (DSA) against HLA and non-HLA antigens in the glomeruli and the tubulointerstitium, which together with high interferon gamma (IFNɣ) and tumor necrosis factor-alpha (TNFα), trigger graft injury. Unfortunately, the mechanisms governing cell-specific injury in AMR remain unclear. We studied 30 for-cause kidney biopsies with early AMR, acute cellular rejection or acute tubular necrosis (‘non-AMR’). We laser-captured microdissected glomeruli and tubulointerstitium and subjected them to unbiased proteome analysis. 120/2026 glomerular and 180/2399 tubulointerstitial proteins were significantly differentially expressed in AMR vs. non-AMR biopsies (p<0.05). Basement membrane and extracellular matrix (ECM) proteins were significantly decreased in both AMR compartments. We verified decreased glomerular and tubulointerstitial LAMC1 expression, and decreased glomerular NPHS1 and PTPRO expression in AMR. Cathepsin-V (CTSV) was predicted to cleave ECM-proteins in the AMR glomeruli, and CTSL, CTSS and LGMN in the tubulointerstitium. We identified galectin-1, an immunomodulatory protein upregulated in AMR glomeruli and linked to the ECM. Anti-HLA class-I antibodies significantly increased CTSV expression, and galectin-1 expression and secretion, in human glomerular endothelial cells. We also studied glutathione S-transferase omega-1 (GSTO1), an ECM-modifying enzyme, increased in the AMR tubulointerstitium. GSTO1 expression was significantly increased in TNFα-treated proximal tubular epithelial cells. IFNɣ and TNFα significantly increased CTSS and LGMN expression in these cells. Basement membranes are often remodeled in chronic AMR, and we demonstrated that this remodeling begins early in glomeruli and tubulointerstitium. Targeting ECM-remodeling in AMR may represent a new therapeutic opportunity.

2020-09-22 | PXD017580 | Pride

TurboID reveals the interaction network of VESICLE-INDUCING PROTEIN IN PLASTIDS (VIPPs) in Chlamydomonas reinhardtii

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Comparative analysis of the human and mouse placental transcriptome and proteome

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