ELECTRON MICROSCOPY OF NETWORK STRUCTURES IN THERMALLY‐INDUCED GLOBULAR PROTEIN GELS

Abstract

Thin sections of heat-set protein gels formed from bovine serum albumin, insulin, lysozyme, ribonuclease, and alpha-chymotrypsin, have been studied by transmission electron microscopy. Micrographs have been interpreted as showing protein networks with strands between one and two times as thick as the native protein diameters. Considerable differences in the persistence characteristics, and frequencies of cross-linking, of the strands are observed, and there are variations in network homogeneity over long distances which correlate well with changes in gel opacity caused by alterations in pH and ionic strength. Evidence that arte facts are unlikely to have influenced these interpretations has been obtained in the BSA case in particular, by studying the aggregation process in solution, using alternative microscope approaches such as heavy-metal shadowing and negative staining. Assuming that artefacts are absent, gel section micrographs have been simulated by a computer procedure, and the results suggest that, in most cases, the simplest interpretation of the microscope data is in terms of a “string of beads” model for the aggregation process, involving only moderately unfolded, and still globular, protein molecules. Other structural interpretations cannot be ruled out, however, as the degree of protein unfolding, and the exact mode of incorporation of the monomers into the network filaments, cannot be established by the microscope technique alone.