Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2022 Mar 2;154(4):e202112965. doi: 10.1085/jgp.202112965

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2022 Rohaim et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms/). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 International license, as described at https://creativecommons.org/licenses/by-nc-sa/4.0/).

PMC Copyright notice

Figure 2. — Expression, purification, and structural determination of the sFab–KcsA complex. (A) An affinity pulldown SDS-PAGE showing the association between KcsA and sFab with (left) and without (right) β-mercaptoethanol (BME). Lane 1: sFab, lane 2–6: sFab–KcsA complex; excess fab is shown in lane 5 and 6 from SEC. (B) Comparison of the analytical size-exclusion chromatograms for the apo (orange) and complex form of KcsA with sFab (blue), with ∼2 ml shift of the elution volume to the left upon complex formation. (C) Crystals of the sFab–KcsA complex visualized under visible (left) and UV (right) light microscope obtained at room temperature after 24 h. (D) The crystal structure of sFab–KcsA where the sFab (magenta) binds at the turret loop of KcsA (green), showing two diagonally opposing monomers for clarity (PDB accession no. 7MDJ). (E) Crystal lattice packing of the sFab–KcsA, structural analysis of the sFab (magenta), and wtFab (gold) showing the positional deviation around the constant domain. (F) Comparison of the interaction between Fabs in crystallographic symmetry mates (Fab^cryst, in orange) showing the asymmetric unit of sFab (left) and wtFab (right). (G) Comparison of the crystal lattice from the new sFab–KcsA on the left with that from wtFab–KcsA (PDB accession no. 1K4C) on the right.