Open AccessThe Folding and Assembly Pathway of Tumour Necrosis Factor TNFα, a Globular Trimeric Protein
Author(s) -
Hlodan Roman,
Pain Roger H.
Publication year - 1995
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1995.0381e.x
Subject(s) - globular protein , tumor necrosis factor alpha , folding (dsp implementation) , protein folding , microbiology and biotechnology , chemistry , biophysics , biology , immunology , crystallography , engineering , structural engineering
The mechanisms of folding and assembly of the globular, trimeric protein tumour necrosis factor‐α (TNF) were studied by chemical cross‐linking. This revealed the rapid accumulation of a dimeric intermediate. Under the conditions of renaturation used, formation of the trimer is complete within six minutes. The kinetics of change of intrinsic and 8‐anilino‐1‐naphthalene sulfonic acid fluorescence are first order and, combined with the kinetics of association, reveal the presence of folding steps both before and subsequent to formation of the trimer. Results from gel exclusion chromatography and kinetics, together with the existence of an acid‐induced molten globule, support the conclusion that TNF folds and assembles through a trimeric molten globule.