Immunoblot studies of the IGF-related acid-labile subunit

J Clin Endocrinol Metab. 1994 Dec;79(6):1883-6. doi: 10.1210/jcem.79.6.7527417.

Abstract

Insulin-like growth factors I and II (IGF-I and II) are present in serum primarily within a ternary complex consisting of IGF, IGF-binding protein-3 (IGF-3) and acid-labile subunit (ALS). Relatively little is known about ALS as compared to the other components of the complex. We report immunoblot studies of ALS using a new rabbit antiserum to human ALS1-34. The antiserum shows high specificity for ALS, labelling only the intact 82-88 kDa doublet in whole serum. Treatment with endoglycosidase-F leads to only a partial deglycosylation of ALS in whole serum, while purified ALS is reduced to M(r) approximately 58 kDa. Acidification of both whole serum and purified ALS leads to a complete loss of ALS ability to bind to cross-linked IGFBP-3:[125I]IGF-II tracer; however, immunoblot studies show no change in the apparent M(r) of the major ALS band. Immunoblot studies of human serum shows that intact ALS is decreased in growth-hormone (GH) deficiency, increases with GH treatment, is elevated in GH excess and is unchanged with IGF-I treatment. These data provide new information regarding the characteristics of ALS and demonstrate the research utility of a highly-specific antiserum for this protein.

MeSH terms

  • Acromegaly / blood
  • Animals
  • Antibody Specificity
  • Carrier Proteins / chemistry
  • Carrier Proteins / immunology
  • Carrier Proteins / metabolism*
  • Female
  • Glycoproteins / chemistry
  • Glycoproteins / immunology
  • Glycoproteins / metabolism*
  • Glycosylation
  • Growth Hormone / deficiency
  • Growth Hormone / pharmacology
  • Humans
  • Hydrogen-Ion Concentration
  • Immune Sera / immunology
  • Immunoblotting*
  • Insulin-Like Growth Factor Binding Proteins
  • Insulin-Like Growth Factor II / metabolism
  • Macromolecular Substances
  • Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / metabolism
  • Molecular Weight
  • Postmenopause / blood
  • Pregnancy

Substances

  • Carrier Proteins
  • Glycoproteins
  • Immune Sera
  • Insulin-Like Growth Factor Binding Proteins
  • Macromolecular Substances
  • insulin-like growth factor binding protein, acid labile subunit
  • Insulin-Like Growth Factor II
  • Growth Hormone
  • Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase