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. Author manuscript; available in PMC: 2022 Jan 4.
Published in final edited form as: J Mol Biol. 2020 Nov 4;432(24):166698. doi: 10.1016/j.jmb.2020.10.032

Fig. 6. DNA-bound PCNA binds one molecule of LigI.

Fig. 6.

Gel filtration to assess LigI-PCNA-DNA complex monodispersity and stoichiometry a. An equimolar mixture of LigI, PCNA, and non-ligatable nicked DNA eluted as a single peak (brown trace). A mixture containing excess LigI produced a second, smaller peak (purple trace) that corresponds to LigI alone. b. SEC-MALS determined correct molar masses of PCNA (green trace), LigI-DNA (blue trace), LigI-PCNA (black trace) and LigI-PCNA-DNA (red trace), confirming equimolar stoichiometry. c. SDS-PAGE (left) and agarose (right) gels identifying LigI, PCNA and DNA within the peak gel filtration fractions.