Fig. 2

Biochemical characterization of SMCHD1 N-terminal constructs. a Domain organization of SMCHD1 N-terminal ATPase module colored as in Fig. 1. b A shift from monomer to dimer in the presence of 5 mM Mg²⁺/ATP is shown for the 24–580 construct. Molecular weights above the peaks are estimates based on elution volumes of standards and are given in kDa. The theoretical molecular weights for the constructs are as follows: 24–406 (44.2 kDa), 110–580 (54.7 kDa), and 24–580 (64.5 kDa). c Dimerization by chemical crosslinking of SMCHD1 constructs (with glutaraldehyde) in the absence (−) and presence (+) of 5 mM Mg²⁺/ATP demonstrates substantial dimerization only in the 24–580 construct (129 kDa). d Radiometric ATPase assay in the three SMCHD1 constructs demonstrates comparable activity for 110–580 and 24–580 but minimal activity for 24–406. Measurements were taken at times 0, 5, 10, 30, 60, and 120 min with at least two biological replicates and are reported as mean ± sd. (Where not obvious, the sd is included in the symbol.)