Skip to main content
Journal cover image

Electron microscopy studies of alpha 2-macroglobulin subunit association after limited reduction with dithiothreitol.

Publication ,  Journal Article
Gonias, SL; Marshall, LB; Figler, NL; Hussaini, IM; Moncino, MD; Pizzo, SV
Published in: Arch Biochem Biophys
April 1993

The subunits of human alpha 2-macroglobulin (alpha 2M) were dissociated by treatment with reductant (0.5 mM dithiothreitol) under mild conditions. Intact tetramers, half-molecules (subunit dimers), and monomers were identified by chromatography on Superose-6. These products were not in rapidly reversible equilibrium since purified half-molecules were completely stable for up to 18 h. Monomers slowly associated to form some half-molecules in the same time period. Negatively stained preparations of monomers and half-molecules demonstrated significant heterogeneity by electron microscopy. This heterogeneity probably reflected structural differences as well as variation in projection and staining. After reaction with methylamine or proteinase, half-molecules and purified monomers reassociated. The principal product was an intact tetramer displaying an "H-like" image which was visually indistinguishable from the structure of intact tetrameric alpha 2M after conformational change. Incompletely reassociated alpha 2M species were also identified after performing chromatography to increase the fraction of these products. Images resembling one-half of the intact tetrameric alpha 2M structure (epsilon-image) or three-quarters of the intact tetramer (chair-image) were observed. These studies demonstrate that reassociation of reductant-dissociated alpha 2M subunits occurs by the formation of appropriate interactions, so that the resulting tetramers are equivalent to nontreated alpha 2M-trypsin. Unlike native alpha 2M, the structure of conformationally transformed alpha 2M is relatively insensitive to the loss of interchain disulfide bonds. We propose that reassembly of alpha 2M subunits occurs by the binding of two half-molecules or by the addition of individual subunits to half-molecules or trimers.

Duke Scholars

Published In

Arch Biochem Biophys

DOI

ISSN

0003-9861

Publication Date

April 1993

Volume

302

Issue

1

Start / End Page

42 / 48

Location

United States

Related Subject Headings

  • alpha-Macroglobulins
  • Trypsin
  • Microscopy, Electron
  • Methylamines
  • Macromolecular Substances
  • Humans
  • Dithiothreitol
  • Chromatography, Gel
  • Biochemistry & Molecular Biology
  • 3101 Biochemistry and cell biology
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Gonias, S. L., Marshall, L. B., Figler, N. L., Hussaini, I. M., Moncino, M. D., & Pizzo, S. V. (1993). Electron microscopy studies of alpha 2-macroglobulin subunit association after limited reduction with dithiothreitol. Arch Biochem Biophys, 302(1), 42–48. https://doi.org/10.1006/abbi.1993.1178
Gonias, S. L., L. B. Marshall, N. L. Figler, I. M. Hussaini, M. D. Moncino, and S. V. Pizzo. “Electron microscopy studies of alpha 2-macroglobulin subunit association after limited reduction with dithiothreitol.Arch Biochem Biophys 302, no. 1 (April 1993): 42–48. https://doi.org/10.1006/abbi.1993.1178.
Gonias SL, Marshall LB, Figler NL, Hussaini IM, Moncino MD, Pizzo SV. Electron microscopy studies of alpha 2-macroglobulin subunit association after limited reduction with dithiothreitol. Arch Biochem Biophys. 1993 Apr;302(1):42–8.
Gonias, S. L., et al. “Electron microscopy studies of alpha 2-macroglobulin subunit association after limited reduction with dithiothreitol.Arch Biochem Biophys, vol. 302, no. 1, Apr. 1993, pp. 42–48. Pubmed, doi:10.1006/abbi.1993.1178.
Gonias SL, Marshall LB, Figler NL, Hussaini IM, Moncino MD, Pizzo SV. Electron microscopy studies of alpha 2-macroglobulin subunit association after limited reduction with dithiothreitol. Arch Biochem Biophys. 1993 Apr;302(1):42–48.
Journal cover image

Published In

Arch Biochem Biophys

DOI

ISSN

0003-9861

Publication Date

April 1993

Volume

302

Issue

1

Start / End Page

42 / 48

Location

United States

Related Subject Headings

  • alpha-Macroglobulins
  • Trypsin
  • Microscopy, Electron
  • Methylamines
  • Macromolecular Substances
  • Humans
  • Dithiothreitol
  • Chromatography, Gel
  • Biochemistry & Molecular Biology
  • 3101 Biochemistry and cell biology