The Sec1–Munc18 protein VPS33B forms a uniquely bidirectional complex with VPS16B

Richard J.Y. Liu; Yusef Al-Molieh; Shao Z. Chen; Marko Drobac; Denisa Urban; Chang H. Chen; Helen H.Y. Yao; Ryan S.Q. Geng; Ling Li; Fred G. Pluthero; Samir Benlekbir; John L. Rubinstein; Walter H.A. Kahr

doi:10.1016/j.jbc.2023.104718

Figure 7

An external file that holds a picture, illustration, etc.
Object name is gr7.jpg

Truncated VPS16B forms a stable complex with VPS33B.A, Coomassie blue–stained SDS-PAGE gel comparing purified VPS33B–VPS16B complex containing wildtype or N-terminal truncated VPS16B. B, SDS-PAGE gel of complex containing VPS33B and amino acids 143 to 316 of VPS16B. C, BN-PAGE gel of VPS33B in complex with amino acids 143 to 493 of VPS16B (arrowhead); truncation does not appear to disrupt complex formation. D, BN-PAGE gel of VPS33B in complex with amino acids 143 to 316 of VPS16B (arrowhead); truncation does not appear to disrupt complex formation. E, 2D class averages from negative-staining EM of crosslinked VPS33B–VPS16B(143–493) complex showing a two-lobed structure similar to full-length complex (Fig. 5B), indicating the stable association of subunits when N terminus of VPS16B is absent. Classes were generated from 308 particles. F, 2D class averages from negative-staining EM of crosslinked VPS33B–VPS16B(143–316) complex showing a two-lobed structure similar to full-length complex, indicating the stable association of subunits when N- and C-terminal regions of VPS16B are absent. Classes were generated from 424 particles. The scale bars represent 10 nm. VPS, vacuolar protein sorting protein.