Characterization of mutated transforming growth factor-beta s which possess unique biological properties

Biochemistry. 1994 Oct 11;33(40):12298-304. doi: 10.1021/bi00206a037.

Abstract

Transforming growth factor-beta (TGF-beta) is a potent regulator of cell growth and differentiation. On the basis of the crystal structure of TGF-beta 2, we have designed and synthesized two mutant TGF-beta s, TGF-beta 1 (71 Trp) and TGF-beta 1 (delta 69-73). Although both of these molecules inhibited the growth of Mv1Lu mink lung epithelial cells and LS1034 colorectal cancer cells, which are affected equally by TGF-beta 1 and TGF-beta 2, TGF-beta 1 (delta 69-73) was much less potent than TGF-beta 1 or TGF-beta 1 (71 Trp) at inhibiting the growth of LS513 colorectal cancer cells which are growth-inhibited by TGF-beta 1 but not TGF-beta 2. Both TGF-beta 1 (71 Trp) and TGF-beta 1 (delta 69-73) increased levels of mRNAs for fibronectin and plasminogen activator inhibitor with Mv1Lu cells, whereas only TGF-beta 1 (71 Trp) and not TGF-beta 1 (delta 69-73) up-regulated the mRNA level of carcinoembryonic antigen in LS513 cells. The expression level of carcinoembryonic antigen mRNA in LS1034 cells was not altered by either wild-type or mutant TGF-beta s. Receptor labeling experiments demonstrated that TGF-beta 1 (71 Trp) bound with high affinity to the cell-surface receptors of Mv1Lu, LS1034, and LS513 cells while TGF-beta 1 (delta 69-73) bound effectively to the receptors of Mv1Lu and LS1034 cells but much less to the receptors on LS513 cells.(ABSTRACT TRUNCATED AT 250 WORDS)

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Binding Sites
  • Blotting, Northern
  • CHO Cells
  • Carcinoembryonic Antigen / genetics
  • Cell Division / drug effects
  • Cell Line
  • Colorectal Neoplasms
  • Cricetinae
  • Cross-Linking Reagents
  • Crystallography, X-Ray
  • Electrophoresis, Polyacrylamide Gel
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / drug effects
  • Feasibility Studies
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • Mink
  • Molecular Sequence Data
  • Mutation / genetics*
  • RNA, Messenger / metabolism
  • Transfection
  • Transforming Growth Factor beta / chemistry*
  • Transforming Growth Factor beta / genetics
  • Transforming Growth Factor beta / pharmacology
  • Tumor Cells, Cultured
  • Umbilical Veins / metabolism
  • Up-Regulation

Substances

  • Carcinoembryonic Antigen
  • Cross-Linking Reagents
  • RNA, Messenger
  • Transforming Growth Factor beta