PMID- 1371050
OWN - NLM
STAT- MEDLINE
DCOM- 19920311
LR  - 20190501
IS  - 0264-6021 (Print)
IS  - 1470-8728 (Electronic)
IS  - 0264-6021 (Linking)
VI  - 281 ( Pt 2)
IP  - Pt 2
DP  - 1992 Jan 15
TI  - Binding of transforming growth factor-beta 1 to methylamine-modified alpha 
      2-macroglobulin and to binary and ternary alpha 2-macroglobulin-proteinase 
      complexes.
PG  - 569-75
AB  - The binding of 125I-labelled transforming growth factor-beta 1 (TGF-beta 1) to 
      human alpha 2-macroglobulin (alpha 2M) was studied by native PAGE and 
      autoradiography. TGF-beta 1 bound preferentially to alpha 2M-methylamine and 
      minimally, if at all, to native alpha 2M. Preparations of alpha 2M-proteinase 
      complex were generated by incubating a standard concentration of alpha 2M (0.4 
      microM) with different concentrations of trypsin, chymotrypsin or neutrophil 
      elastase (0.04-2.0 microM). The 125I-TGF-beta 1-binding activity depended on the 
      initial ratio of active proteinase to alpha 2M, or r value, used to form the 
      alpha 2M-proteinase complex. With all three proteinases, r values of 2 or greater 
      yielded preparations with unchanged or decreased TGF-beta 1-binding activity 
      relative to native alpha 2M. By contrast, r values near 1 yielded preparations 
      with significantly increased TGF-beta 1-binding activity. The results of 
      [3H]thymidine-incorporation studies performed in mouse keratinocytes were 
      consistent with the 125I-TGF-beta-binding experiments. alpha 2M-trypsin and alpha 
      2M-chymotrypsin prepared at an r value of 1.0 counteracted the activity of 
      TGF-beta 1, whereas the equivalent complexes prepared at an r value of 3.0 had no 
      effect. As determined by SDS/PAGE, 125I-TGF-beta 1 binding to alpha 
      2M-methylamine was at least 80% non-covalent. Reaction of alpha 2M-methylamine 
      with iodoacetamide or 5,5'-dithiobis-(2-nitrobenzoic acid) decreased the 
      percentage of covalent binding but had no effect on total binding. Neuraminidase 
      treatment had no effect on the binding of 125I-TGF-beta 1 to alpha 
      2M-methylamine. Cleavage of the 'bait regions' in alpha 2M-methylamine by 
      prolonged treatment with trypsin also had no effect. These studies suggest that 
      TGF-beta 1 binding to alpha 2M is enhanced by conformational change in the 
      proteinase inhibitor resulting from reaction with proteinase or amine. If both 
      proteinase-binding sites in a single alpha 2M molecule are occupied, TGF-beta 
      1-binding activity is decreased or perhaps eliminated.
FAU - Hall, S W
AU  - Hall SW
AD  - Department of Pathology, University of Virginia Health Sciences Center, 
      Charlottesville 22908.
FAU - LaMarre, J
AU  - LaMarre J
FAU - Marshall, L B
AU  - Marshall LB
FAU - Hayes, M A
AU  - Hayes MA
FAU - Gonias, S L
AU  - Gonias SL
LA  - eng
GR  - GM 07267/GM/NIGMS NIH HHS/United States
GR  - HL-02272/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - England
TA  - Biochem J
JT  - The Biochemical journal
JID - 2984726R
RN  - 0 (Methylamines)
RN  - 0 (Transforming Growth Factor beta)
RN  - 0 (alpha-Macroglobulins)
RN  - EC 3.2.1.18 (Neuraminidase)
RN  - EC 3.4.- (Endopeptidases)
RN  - EC 3.4.21.1 (Chymotrypsin)
RN  - EC 3.4.21.36 (Pancreatic Elastase)
RN  - EC 3.4.21.4 (Trypsin)
SB  - IM
MH  - Animals
MH  - Cells, Cultured
MH  - Chymotrypsin/metabolism
MH  - Electrophoresis, Polyacrylamide Gel
MH  - Endopeptidases/*metabolism
MH  - Humans
MH  - Keratinocytes/metabolism
MH  - Methylamines/pharmacology
MH  - Mice
MH  - Mice, Inbred BALB C
MH  - Neuraminidase/metabolism
MH  - Pancreatic Elastase/metabolism
MH  - Transforming Growth Factor beta/antagonists & inhibitors/*metabolism
MH  - Trypsin/metabolism
MH  - alpha-Macroglobulins/chemistry/*metabolism
PMC - PMC1130723
EDAT- 1992/01/15 00:00
MHDA- 1992/01/15 00:01
PMCR- 1992/01/15
CRDT- 1992/01/15 00:00
PHST- 1992/01/15 00:00 [pubmed]
PHST- 1992/01/15 00:01 [medline]
PHST- 1992/01/15 00:00 [entrez]
PHST- 1992/01/15 00:00 [pmc-release]
AID - 10.1042/bj2810569 [doi]
PST - ppublish
SO  - Biochem J. 1992 Jan 15;281 ( Pt 2)(Pt 2):569-75. doi: 10.1042/bj2810569.