ATP13A3 is a major component of the enigmatic mammalian polyamine transport system

Polyamines, such as putrescine, spermidine and spermine, are physiologically important polycations, but the transporters responsible for their uptake in mammalian cells remain poorly characterized. Here, we reveal a new component of the mammalian polyamine transport system (PTS) using CHO-MG cells, a widely used model to study alternative polyamine uptake routes and characterize polyamine transport inhibitors for therapy. CHO-MG cells present polyamine uptake deficiency and resistance to a toxic polyamine biosynthesis inhibitor MGBG (methylglyoxal bis- (guanylhydrazone)), but the molecular defects responsible for these cellular characteristics remain unknown. By genome sequencing of CHO-MG cells, we identified mutations in an unexplored gene, ATP13A3, and found disturbed mRNA and protein expression. ATP13A3 encodes for an orphan P5B-ATPase (ATP13A3), a P-type transport ATPase that represents a candidate polyamine transporter. Interestingly, ATP13A3 complemented the putrescine transport deficiency and MGBG resistance of CHO-MG cells, whereas its knockdown in wild-type cells induced a CHO-MG phenotype, demonstrating a decrease in putrescine uptake and MGBG sensitivity. Taken together, our findings identify ATP13A3 as a major component of the mammalian PTS that confers sensitivity to MGBG and that has been previously genetically linked with pulmonary arterial hypertension.

Polyamine transport system-targeted nanometric micelles assembled from epipodophyllotoxin-amphiphiles

Micelle-forming amphiphilic drug conjugates were synthesized starting from a biologically active epipodophyllotoxin derivative which was covalently inserted in between a hydrophilic targeting spermine unit, and a hydrophobic stearyl chain.

Spermine-NBD as Fluorescent Probe for Studies of Polyamine Transport System in Leishmania Donovani

This study describes the synthesis of fluorescent probes as potential substrates for the polyamine transport system (PTS) of <i>Leishmania donovani</i>. A competitive radioassay was used to determine the most efficient probe. We observed that the conjugate spermine-nitrobenzofurazan (Spm-NBD) was able to compete with [³H]-spermidine in <i>L. donovani</i> at a relevant IC₅₀ of 60 µM. <p><br></p>

Spermine-NBD as Fluorescent Probe for Studies of Polyamine Transport System in Leishmania Donovani

This study describes the synthesis of fluorescent probes as potential substrates for the polyamine transport system (PTS) of <i>Leishmania donovani</i>. A competitive radioassay was used to determine the most efficient probe. We observed that the conjugate spermine-nitrobenzofurazan (Spm-NBD) was able to compete with [³H]-spermidine in <i>L. donovani</i> at a relevant IC₅₀ of 60 µM. <p><br></p>