PAINT places BOLA3 protein maturation and mitochondrial-matrix assertions at PANTHER:PTN001763701.

BOLA1 counteracts glutathione-depletion effects on mitochondrial thiol redox potential and morphology. The Introduction separately places BOLA3 in mitochondrial Fe-S biogenesis; the BOLA1 experiments do not establish a BOLA3 redox function.

Affinity-enrichment mass spectrometry maps condition-specific mitochondrial protein interactions. Follow-up experiments identify C17orf89 as a complex-I assembly factor and LYRM5-dependent deflavination of electron-transferring flavoprotein.

Yeast genetics places mitochondrial Bol1/Bol3 in late Fe-S protein maturation, with a major LIAS-related phenotype; the authors explicitly leave aspects of the biochemical mechanism unresolved.

Human-protein NMR detects apo and reconstituted holo GLRX5–BOLA3 association. In contrast, size-exclusion chromatography does not show a stable holo heterodimer; CD effects are weak and GSH-dependent, and BOLA3 does not stabilize the cluster against dithionite. These results distinguish detectable interaction from kinetic stability under the tested conditions.

Microscale thermophoresis reports direct BOLA3–NFU1 interaction, preferentially with holo-NFU1.

Bol1/Bol3 deletion did not activate the yeast iron regulon; Bol2-dependent effects and indirect metabolic crosstalk are distinguished from a direct mitochondrial Bol role in iron regulation.

Yeast Bol3 loss mildly reduces succinate dehydrogenase activity; combined Bol1/Bol3 loss produces stronger Fe-S-enzyme and lipoylation defects rescued by either Bol protein. The Discussion relates this specificity to LIAS and respiratory-complex I/II deficiencies in BOLA3-deficient patients.

HuRI maps approximately 53,000 human binary protein interactions and links the network to tissue-context and Mendelian-disease analyses.

Affinity-purification mass spectrometry in 293T and HCT116 cells reveals shared core protein complexes and extensive cell-context-dependent interaction remodeling.

C59Y maintains the BOLA3 fold and GLRX5 association while altering interaction geometry and cluster coordination; it does not abolish cluster binding.

The human reconstitution/NMR comparison measures less cluster-bound NFU1 with C59Y than wild-type BOLA3–GLRX5. The proposed explanation of one compound-heterozygous patient’s recovery remains a mechanistic interpretation, not a demonstrated developmental rescue mechanism.

Quantitative human-cell mitochondrial proteomics defines a high-confidence set of more than 1,100 proteins and measures broad variation in abundance and turnover, with additional analyses of import and protein-complex assembly.