Analysis of the Cys82Arg mutation in follicle-stimulating hormone beta (FSHbeta) using a novel FSH expression vector

Fertil Steril. 2003 Feb;79(2):379-85. doi: 10.1016/s0015-0282(02)04660-5.

Abstract

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.

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

Setting: Tertiary medical center setting.

Patient(s): DNA sequence of the FSHbeta gene and clinical description from a patient with isolated FSH deficiency.

Intervention(s): 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.

Main outcome measure(s): Immunoreactive and bioactive FSH levels from the CHO cellular media.

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

Conclusion(s): 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.

Publication types

  • Case Reports
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adult
  • Amino Acid Substitution
  • Animals
  • Arginine
  • CHO Cells
  • Cricetinae
  • Cysteine
  • DNA / genetics*
  • Follicle Stimulating Hormone / deficiency*
  • Follicle Stimulating Hormone, beta Subunit / chemistry
  • Follicle Stimulating Hormone, beta Subunit / genetics*
  • Genetic Vectors
  • Humans
  • Male
  • Mutagenesis, Site-Directed
  • Oligospermia / genetics
  • Polymerase Chain Reaction
  • Recombinant Proteins / chemistry
  • Transfection

Substances

  • Follicle Stimulating Hormone, beta Subunit
  • Recombinant Proteins
  • Follicle Stimulating Hormone
  • DNA
  • Arginine
  • Cysteine