title: Expression of biologically active human follitropin in Chinese hamster ovary cells.
abstract: To study the structure-function relationships of follitropin (FSH), we expressed the hormone in a heterologous cell system. A genomic clone bearing a 3.7-kilobase FSH beta insert containing the entire coding sequence was transfected alone or together with the alpha subunit gene into Chinese hamster ovary cells and stable lines expressing either FSH beta or FSH dimer were selected. Pulse-chase experiments revealed that, when transfected alone FSH beta was very slowly secreted similar to lutropin beta and thyrotropin beta but unlike choriogonadotropin beta which is efficiently secreted. However, cotransfection of the FSH beta and alpha subunit genes resulted in "rescue" of the beta subunit and rapid secretion of dimer. These data support the hypothesis that the glycoprotein hormones of pituitary origin have determinants for secretion that differ from those on the placental hormone, choriogonadotropin. Recombinant FSH stimulated steroidogenesis comparable to purified human FSH isolated from pituitaries in an in vitro rat granulosa cell assay and appears more homogeneous by chromatofocusing. Human FSH produced by this cell line provides a source of bioactive FSH for experimental and clinical use.
authors.affiliation: Department of Pharmacology, Washington University School of Medicine, St. Louis, Missouri 63110.
authors.forename: J L
authors.initials: JL
authors.lastname: Keene
authors.forename: M M
authors.initials: MM
authors.lastname: Matzuk
authors.forename: T
authors.initials: T
authors.lastname: Otani
authors.forename: B C
authors.initials: BC
authors.lastname: Fauser
authors.forename: A B
authors.initials: AB
authors.lastname: Galway
authors.forename: A J
authors.initials: AJ
authors.lastname: Hsueh
authors.forename: I
authors.initials: I
authors.lastname: Boime
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D005260:Female; D005640:Follicle Stimulating Hormone / Q000235:genetics* / Q000302:isolation & purification / Q000502:physiology; D005796:Genes; D006801:Humans; D007525:Isoelectric Focusing; D008969:Molecular Sequence Data; D010053:Ovary / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000502:physiology
chemical_list: D011994:Recombinant Proteins; D005640:Follicle Stimulating Hormone