Visualization of membrane-associated folate transport proteins.
Transport of Methotrexate (MTX) into cells, via the "reduced folate" transport system, is a critical factor in the effectiveness of the drug in cancer chemotherapy, and defective transport is one of the principal types of resistance to MTX. Probes capable of detecting membrane-associated folate transport proteins (ftp's) in individual cells are potentially useful for identifying structural and functional domains and for investigating mechanisms of substrate translocation. Polyclonal antibody to highly purified ftp from Lactobacillus casei, in conjunction with a second, gold-labeled antibody, has been used to visualize, via electron microscopy, the protein in Triton-treated membrane fragments and in the membrane and cytoplasm of spheroplasts. To visualize ftp in L1210 cells, the substrate-binding site was first labeled covalently with activated fluorescein-Methotrexate, and the cells were then treated with anti-fluorescein antibody and the gold-labeled antibody.
Pope
L E
LE
Department of Basic and Clinical Research, Research Institute of Scripps Clinic, La Jolla, California 92037.
Fan
J
J
Chang
C M
CM
Minskoff
S A
SA
Vitols
K S
KS
Huennekens
F M
FM
D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism* / Q000648:ultrastructure; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure; D058973:Folate Receptors, GPI-Anchored; D005492:Folic Acid / Q000378:metabolism*; D007150:Immunohistochemistry / Q000379:methods; D007780:Lacticaseibacillus casei / Q000378:metabolism; D007939:Leukemia L1210 / Q000378:metabolism; D051379:Mice; D008854:Microscopy, Electron / Q000379:methods; D011956:Receptors, Cell Surface
D002352:Carrier Proteins; D058973:Folate Receptors, GPI-Anchored; D011956:Receptors, Cell Surface; D005492:Folic Acid