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Cross‐Linked 7S Nerve Growth Factor Is Biologically Inactive

Abstract: The 7S nerve growth factor complex was cross‐linked with dimethylsuberimidate in a pH jump reaction and the cross‐linked product of highest molecular weight was separated from non‐cross‐linked subunits by gel filtration at acid pH. The amount of the cross‐linked 7S nerve growth factor generated was reduced by exposure of the complex to pH 9.5, where it dissociates. The cross‐linked 7S nerve growth factor migrated as a single species on electrophoresis under denaturing and reducing conditions and had a molecular weight, 132,000, identical to that of 7S nerve growth factor. All three subunits of the complex were released on deamidination. The biological activity of the cross‐linked 7S nerve growth factor complex was less than 0.3% that of native βNGF, suggesting that the biological activity of the latter is completely inhibited by interactions with other subunits in the complex.