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Dimethyl suberimidate is a bifunctional chemical reagent used for protein cross-linking. Its two imidoester ends can react with amino groups to form amidine linkages, allowing a single reagent molecule to connect two accessible protein groups.^[ref]

In 1970, G. E. Davies and G. R. Stark used the reagent to investigate subunit organization in oligomeric proteins. After cross-linking, detergent gel electrophoresis resolved species with apparent masses corresponding to integral multiples of the individual subunit mass.^[ref]

For an illustrative $20\mathrm{kDa}$ subunit, linked pairs and triples would appear near $40$ and $60\mathrm{kDa}$ before accounting for linker mass and electrophoretic behaviour. A covalent bridge can preserve an association through conditions that disrupt the original noncovalent interactions.

The observation establishes chemical proximity under the reaction conditions. It does not automatically locate a unique native interface or prove that every detected pair is a stable complex in a living cell. Reagent accessibility, possible links within one chain and collisions between molecules all affect the products. Comparing treated and untreated preparations, and identifying the protein species, makes the structural inference more informative.

G. E. Davies coauthored a 1970 method for investigating protein quaternary structure through protein cross-linking. With G. R. Stark, G. E. Davies used dimethyl suberimidate, a reagent with two reactive ends, to join nearby protein subunits before analysing the resulting products.^[ref]

SDS-PAGE resolved products with masses equal to multiples of the individual protein subunit mass. A series of linked products could therefore reveal an assembly's subunit organization after treatment that would otherwise separate its components.^[ref]

For an invented three-subunit assembly with $20\mathrm{kDa}$ per subunit, partial linking could produce species near $20$, $40$ and $60\mathrm{kDa}$. Seeing only the middle species would not establish that the original assembly contained two subunits: the completeness and selectivity of the linking reaction matter.

G. R. Stark coauthored, with G. E. Davies, a 1970 experiment using protein cross-linking to investigate the organization of oligomeric proteins. The method connected a chemical intervention to a measurable series of protein subunit masses.^[ref]

After reaction with dimethyl suberimidate, detergent gel electrophoresis separated individual chains and covalently linked combinations. The experiments included assemblies with identical subunits and assemblies containing different kinds of subunits; the latter showed unequal chemical reactivities.^[ref]

The unequal-reactivity observation gives an important interpretive control. Failure to observe a linked pair need not mean that its members were far apart in the native protein: reactive groups may have been inaccessible or poorly reactive. Chemical proximity measurements must therefore be interpreted together with the chemistry used to record them.