Physical and biological characterization of a growth-inhibitory activity purified from the neuroepithelioma cell line A673

Biochem J. 1995 Jan 1;305 ( Pt 1)(Pt 1):87-92. doi: 10.1042/bj3050087.

Abstract

Epithelial- and haematopoietic-cell growth-inhibitory activities have been identified in the conditioned medium of the human peripheral neuroepithelioma cell line A673. An A673-cell-derived growth-inhibitory activity was previously fractionated into two distinct components which inhibited the proliferation of human carcinoma and leukaemia cells in culture. One inhibitory activity was shown to comprise interleukin-1 alpha (IL-1 alpha). Here, we have purified to homogeneity a distinct activity which inhibited the growth of the epithelial cells in vitro. Using a combination of protein-sequence analysis and mass spectrometry, we demonstrated that biological activity can be assigned to a dimeric protein with a molecular mass of 25,576 (+/- 4) Da and an N-terminal sequence identical with that of transforming growth factor-beta 1 (TGF-beta 1). Further characterization of the growth inhibitor with TGF-beta-isoform-specific antibodies showed that > 90% of the bioactivity consists of TGF-beta 1 and not TGF-beta 2 or TGF-beta 3. Although A673 cells were growth-inhibited by exogenous TGF-beta 1, we showed that TGF-beta 1 in A673-cell-conditioned media was present in the latent, biologically inactive, form which did not act as an autocrine growth modulator of A673 cells in vitro.

Publication types

  • Comparative Study
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Antibodies / pharmacology
  • Antibody Specificity
  • Cell Division / drug effects
  • Cell Division / physiology
  • Chemical Phenomena
  • Chemistry, Physical
  • Culture Media
  • Growth Substances / chemistry
  • Growth Substances / isolation & purification*
  • Growth Substances / physiology*
  • Humans
  • Molecular Sequence Data
  • Neuroectodermal Tumors, Primitive, Peripheral / chemistry*
  • Neuroectodermal Tumors, Primitive, Peripheral / pathology*
  • Neutralization Tests
  • Sequence Homology, Amino Acid
  • Transforming Growth Factor beta / analysis
  • Transforming Growth Factor beta / immunology
  • Tumor Cells, Cultured

Substances

  • Antibodies
  • Culture Media
  • Growth Substances
  • Transforming Growth Factor beta