Analysis of the Cys82Arg mutation in follicle-stimulating hormone beta (FSHbeta) using a novel FSH expression vector.
OBJECTIVE
To determine the effect of the Cys82Arg FSHbeta mutation from a patient with isolated FSH deficiency upon follicle-stimulating hormone (FSH) levels in vitro.


METHODS
In vitro analysis of the Cys82Arg mutation and comparison with the phenotype.


METHODS
Tertiary medical center setting.


METHODS
DNA sequence of the FSHbeta gene and clinical description from a patient with isolated FSH deficiency.


METHODS
Construction of a new vector containing the cDNAs for the alpha-subunit and beta-subunit of FSH (palphaFSHbeta) followed by mutagenesis and transfection into Chinese hamster ovary cells.


METHODS
Immunoreactive and bioactive FSH levels from the CHO cellular media.


RESULTS
Although expression of both subunits was present, both immunoreactive and bioactive FSH levels were unmeasurable from cellular media containing the mutation versus wild type.


CONCLUSIONS
The Cys82Arg mutation in a male with normal puberty and azoospermia results in profound deficiency of FSH in vitro, thereby confirming the molecular basis of hypogonadism in this patient and documenting the importance of the Cys residue at position 82 of the FSHbeta subunit.
Clark
Andrew D
AD
Section of Reproductive Endocrinology, Infertility and Genetics, Department of Obstetrics and Gynecology, Medical College of Georgia, Augusta, Georgia 30912, USA.
Layman
Lawrence C
LC
D000328:Adult; D019943:Amino Acid Substitution; D000818:Animals; D001120:Arginine; D016466:CHO Cells; D006224:Cricetinae; D003545:Cysteine; D004247:DNA / Q000235:genetics*; D005640:Follicle Stimulating Hormone / Q000172:deficiency*; D037201:Follicle Stimulating Hormone, beta Subunit / Q000737:chemistry / Q000235:genetics*; D005822:Genetic Vectors; D006801:Humans; D008297:Male; D016297:Mutagenesis, Site-Directed; D009845:Oligospermia / Q000235:genetics; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000737:chemistry; D014162:Transfection
D037201:Follicle Stimulating Hormone, beta Subunit; D011994:Recombinant Proteins; D005640:Follicle Stimulating Hormone; D004247:DNA; D001120:Arginine; D003545:Cysteine