The peroxisomal protein PXN encoded by the Arabidopsis gene At2g39970 has very recently been found to transport NAD(+), NADH, AMP and ADP. In this work we have reinvestigated the substrate specificity and the transport properties of PXN by using a wide range of potential substrates. Heterologous expression in bacteria followed by purification, reconstitution in liposomes, and uptake and efflux experiments revealed that PNX transports coenzyme A (CoA), dephospho-CoA, acetyl-CoA and adenosine 3', 5'-phosphate (PAP), besides NAD(+), NADH, AMP and ADP. PXN catalyzed fast counter-exchange of substrates and much slower uniport and was strongly inhibited by pyridoxal 5'-phosphate, bathophenanthroline and tannic acid. Transport was saturable with a submillimolar affinity for NAD(+), CoA and other substrates. The physiological role of PXN is probably to provide the peroxisomes with the essential coenzymes NAD(+) and CoA.
The peroxisomal NAD(+) carrier of Arabidopsis thaliana transports coenzyme A and its derivatives.
AGRIMI, GENNARO;PIERRI, CIRO LEONARDO;
2012-01-01
Abstract
The peroxisomal protein PXN encoded by the Arabidopsis gene At2g39970 has very recently been found to transport NAD(+), NADH, AMP and ADP. In this work we have reinvestigated the substrate specificity and the transport properties of PXN by using a wide range of potential substrates. Heterologous expression in bacteria followed by purification, reconstitution in liposomes, and uptake and efflux experiments revealed that PNX transports coenzyme A (CoA), dephospho-CoA, acetyl-CoA and adenosine 3', 5'-phosphate (PAP), besides NAD(+), NADH, AMP and ADP. PXN catalyzed fast counter-exchange of substrates and much slower uniport and was strongly inhibited by pyridoxal 5'-phosphate, bathophenanthroline and tannic acid. Transport was saturable with a submillimolar affinity for NAD(+), CoA and other substrates. The physiological role of PXN is probably to provide the peroxisomes with the essential coenzymes NAD(+) and CoA.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
