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. 2020 May 11;21(9):3403. doi: 10.3390/ijms21093403

Figure 3.

Figure 3

Hetero-complex formation between hMPC-1 and hMPC-2. (A,B) Analytical size-exclusion gel chromatography (aSEC) assay for identifying homo- and hetero-complexes of hMPCs. The monodisperse peaks of hMPC-1, hMPC-2, and T-hMPC-2 are represented by a dark gray dashed line, dash-double dotted line, and light gray dotted line, respectively. Both peaks of T-hMPC-2/hMPC-1 and T-hMPC-2/hMPC-2 are shown as a black line. The retention volumes of hMPC-1, hMPC-2, T-hMPC-2, T-hMPC-2/hMPC-1, and T-hMPC-2/hMPC-2 were 79.97 mL, 79.69 mL, 77.63 mL, 78.23 mL, and 79.46 mL, respectively. All aSEC analyses were performed using HiLoad Superdex 200 PG equilibrated with buffer containing 50 mM Tris-HCl (pH 7), 300 mM NaCl, and 0.05% (w/v) DDM. Absorbance was monitored at 280 nm. The peak shifting indicates by a red arrow. (C,D) SDS-PAGE analysis of eluted proteins across the retention volume (mL). The peaks containing T-hMPC-2/hMPC-1 (C) and T-hMPC-2/hMPC-2 (D) were isolated by each fraction separately, and protein contents were analyzed by SDS-PAGE. (E,F) Schematic diagram of the proposed process for the homo- and hetero-complexes of hMPC-1 and hMPC-2 based on the experimental setup. The higher strength of binding and dissociation constants about MPCs heterodimerization was highlighted in blue and red arrows, respectively.