Human Mast Cells Secreting Leukotriene C_4 Express the MRP1 Gene-Encoded Conjugate Export Pump

摘要

Mast cells are known to secrete endogenously synthesized leukotriene C_4 (LTC_4), but the identity of the responsible export pump in human mast cells was unknown. The multidrug resistance proteins MRP1 and MRP2 have been identified as primary-active ATP-dependent export pumps for various amphiphilic anions including the glutathione conjugate LTC_4. We therefore studied the expression at the RNA and protein levels of both MRP1 and MRP2 as well as the ATP-dependent LTC_4 transport in the human mast cell line HMC-1. Upon stimulation by 1 μm ionomycin, intact HMC-1 cells generates 26 pmol LTC_4/10~8 cells within 20 min. Transport experiments using inside-out HMC-1 membrane vesicles demonstrated an ATP-dependent LTC_4 transport amounting to 1.4 pmol * (mg protein)~(-1) * min~(-1). Reverse transcription PCR indicated that HMC-1 cells express mRNA of MRP1, but not of MRP2 or MRP3. Cloning and sequencing of the amplified PCR fragment confirmed its identity with the human MRP1 sequence. Immunoblots using antibodies against MRP1 and MRP2 demonstrated that HMC-1 cells contain the MRP1 but not the MRP2 protein. Our results indicate that the 190 kDa integral membrane glycoprotein MRP1 mediates the ATP-dependent export of LTC_4 from human mast cells to the extracellular space.