Characterization of a high-affinity folate receptor in normal and malignant human testicular tissue.
We have characterized the folate receptor in normal and malignant tissue from male gonads. Radioligand binding displayed characteristics typical of other folate receptors. Those included a high-affinity type of binding (K = 10(10M-1)), apparent positive cooperativity changing into non-cooperativity at low receptor concentrations, a tendency to increased binding affinity with decreasing receptor concentrations, a slow dissociation at pH 7.4 becoming rapid at pH 3.5 and inhibition by folates, in particular oxidized forms. The gel filtration profile of Triton X-100 solubilized tissue contained a 25 and 100 kDa peak of radioligand-receptor. The latter peak could represent receptor equipped with a hydrophobic membrane anchor that inserts into Triton X-100 micelles. The concentration of radiolabelled receptor ranged from 0.41 nmol/g protein to 1.68 nmol/g protein in specimens of normal testicular tissue from patients with prostatic carcinomas and from 1.54 nmol/g protein to 3.82 nmol/g protein in testicular tissue from young individuals. Compared to normal testicular tissue the concentration of receptor in seminoma tissue was low (0.38-1.27 nmol/g protein) but showed a higher degree of immunoreactivity in the presence of antibodies against human milk folate binding protein as evidenced by ELISA and immunohistochemistry data. Hence a folate receptor isoform homologous to human milk folate binding protein is apparently expressed in seminomas where the total expression of receptor, however, seems to be lower than in normal testicles.
Holm
J
J
Department of Clinical Chemistry, Central Hospital Herning, Denmark.
Hansen
S I
SI
Høier-Madsen
M
M
Christensen
T B
TB
Nichols
C W
CW
D000368:Aged; D000369:Aged, 80 and over; D002352:Carrier Proteins / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism*; D002850:Chromatography, Gel; D058973:Folate Receptors, GPI-Anchored; D005492:Folic Acid / Q000378:metabolism; D005493:Folic Acid Antagonists / Q000494:pharmacology; D006801:Humans; D007150:Immunohistochemistry; D002955:Leucovorin / Q000494:pharmacology; D008297:Male; D008727:Methotrexate / Q000494:pharmacology; D008970:Molecular Weight; D011956:Receptors, Cell Surface; D012016:Reference Values; D018239:Seminoma / Q000188:drug therapy / Q000378:metabolism*; D013736:Testicular Neoplasms / Q000188:drug therapy / Q000378:metabolism*; D013737:Testis / Q000187:drug effects / Q000378:metabolism*; D014316:Tritium
D002352:Carrier Proteins; D058973:Folate Receptors, GPI-Anchored; D005493:Folic Acid Antagonists; D011956:Receptors, Cell Surface; D014316:Tritium; D005492:Folic Acid; D002955:Leucovorin; D008727:Methotrexate