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

PMC full text:

J Biol Chem. 2023 Jun; 299(6): 104718.

Published online 2023 Apr 14. doi: 10.1016/j.jbc.2023.104718

Copyright/LicenseRequest permission to reuse

Figure 4

An external file that holds a picture, illustration, etc.
Object name is gr4.jpg

Open in a separate window

The VPS33B–VPS16B complex contains two VPS33B and three VPS16B proteins.A, SDS-PAGE gel of HA-VPS33B–HA-VPS16B-3xFLAG complex purified from yeast lysate via FLAG affinity purification and anion exchange chromatography. B, serially diluted samples of complex (total protein shown for each lane) were blotted and probed with monoclonal anti-HA primary antibody and Alexa Fluor 488–conjugated secondary antibody; box indicates 10 ng bands used for quantification (see the Experimental procedures section). C, analysis of fluorescence signal showed 39.1% of total corresponded to HA-VPS33B and 60.9% to HA-VPS16B-3xFLAG, indicating a 2:3 ratio of these proteins in the complex. HA, hemagglutinin tag; VPS, vacuolar protein sorting protein.

Images in this article

Click on the image to see a larger version.