Antibodies against pyridoxal 5′-phosphate and pyridoxamine 5′-phosphate

Córdoba, Félix; González, C.; Rivera, Pablo

doi:10.1016/0304-4165(66)90485-5

Cited by 22 publications

(1 citation statement)

References 7 publications

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“…Serum albumin is the principal protein transporting PLP in the blood [40, 41], and commercially prepared samples generally contain bound PLP cofactor. Also, commercially available anti-conjugated pyridoxal antibodies are raised against pyridoxal-conjugated BSA [42, 43], so they may be expected to be particularly sensitive to that species. The total protein content of the purified mitochondria was determined by measuring the 280 nm absorbance for samples solubilized in 0.6% SDS.…”

Section: Resultsmentioning

confidence: 99%

The Mtm1p carrier and pyridoxal 5′-phosphate cofactor trafficking in yeast mitochondria

Whittaker¹,

Penmatsa²,

Whittaker³

2015

Archives of Biochemistry and Biophysics

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Biochemical communication between the cytoplasmic and mitochondrial subsystems of the cell depends on solute carriers in the mitochondrial inner membrane that transport metabolites between the two compartments. We have expressed and purified a yeast mitochondrial carrier protein (Mtm1p, YGR257cp), originally identified as a manganese ion carrier, for biochemical characterization aimed at resolving its function. High affinity, stoichiometric pyridoxal 5′-phosphate (PLP) cofactor binding was characterized by fluorescence titration and calorimetry, and the biochemical effects of mtm1 gene deletion on yeast mitochondria were investigated. The PLP status of the mitochondrial proteome (the mitochondrial ‘PLP-ome’) was probed by immunoblot analysis of mitochondria isolated from wild type (MTM1+) and knockout (MTM1−) yeast, revealing depletion of mitochondrial PLP in the latter. A direct activity assay of the enzyme catalyzing the first committed step of heme biosynthesis, the PLP-dependent mitochondrial enzyme 5-aminolevulinate synthase, extends these results, providing a specific example of PLP cofactor limitation. Together, these experiments support a role for Mtm1p in mitochondrial PLP trafficking and highlight the link between PLP cofactor transport and iron metabolism, a remarkable illustration of metabolic integration.

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