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. 2021 Jul 15;220(9):e202005214. doi: 10.1083/jcb.202005214

Figure 2.

Figure 2.

Characterization of the TLNRD1 dimer. (A and B) The symmetrical antiparallel TLNRD1 dimer interface mediated by F250. One monomer is shown as a white surface representation, F250 from the opposing monomer (orange) inserts into the pocket on the surface of the other. A salt bridge between R246 and E267’ is shown. (C and D) SEC-MALS analysis of 106 µM TLNRD1-FL WT (black) and F250D (red). (D) Analysis of the molar mass of the major TLNRD1-FL species “a” yields a molecular weight of ∼74 kD. The TLNRD1-F250D yields a molecular weight of ∼37.8 kD, peak “c.” (E) TLNRD1-FL dimerization measured using MST at 25°C with 40% Red LED laser excitation. Unlabeled TLNRD1-FL was titrated into a fixed concentration (50 nM) of fluorescently labeled TLNRD1-FL. Data were analyzed in MO.Affinity Analysis software (v2.1.3) using the law of mass action to generate an apparent equilibrium dimer Kd, Kd,dimer of 80 ± 0.6 nM. F250D Kd,dimer not determined. FNorm, normalized fluorescence; ND, not defined.