+Search query
-Structure paper
Title | Synthetic antibodies against BRIL as universal fiducial marks for single-particle cryoEM structure determination of membrane proteins. |
---|---|
Journal, issue, pages | Nat Commun, Vol. 11, Issue 1, Page 1598, Year 2020 |
Publish date | Mar 27, 2020 |
Authors | Somnath Mukherjee / Satchal K Erramilli / Mark Ammirati / Frances J D Alvarez / Kimberly F Fennell / Michael D Purdy / Blazej M Skrobek / Katarzyna Radziwon / John Coukos / Yanyong Kang / Przemysław Dutka / Xiang Gao / Xiayang Qiu / Mark Yeager / H Eric Xu / Seungil Han / Anthony A Kossiakoff / |
PubMed Abstract | We propose the concept of universal fiducials based on a set of pre-made semi-synthetic antibodies (sABs) generated by customized phage display selections against the fusion protein BRIL, an ...We propose the concept of universal fiducials based on a set of pre-made semi-synthetic antibodies (sABs) generated by customized phage display selections against the fusion protein BRIL, an engineered variant of apocytochrome b562a. These sABs can bind to BRIL fused either into the loops or termini of different GPCRs, ion channels, receptors and transporters without disrupting their structure. A crystal structure of BRIL in complex with an affinity-matured sAB (BAG2) that bound to all systems tested delineates the footprint of interaction. Negative stain and cryoEM data of several examples of BRIL-membrane protein chimera highlight the effectiveness of the sABs as universal fiducial marks. Taken together with a cryoEM structure of sAB bound human nicotinic acetylcholine receptor, this work demonstrates that these anti-BRIL sABs can greatly enhance the particle properties leading to improved cryoEM outcomes, especially for challenging membrane proteins. |
External links | Nat Commun / PubMed:32221310 / PubMed Central |
Methods | EM (single particle) / X-ray diffraction |
Resolution | 1.872 - 3.87 Å |
Structure data | EMDB-20857, PDB-6ur8: EMDB-20863, PDB-6usf: PDB-6cbv: |
Chemicals | ChemComp-GOL: ChemComp-FMT: ChemComp-NA: ChemComp-HOH: ChemComp-NAG: ChemComp-QMR: ChemComp-BMA: |
Source |
|
Keywords | IMMUNE SYSTEM / 4-helix bundle / fusion protein / synthetic antibody / Fab fragment / fusion protein in complex with Fab / MEMBRANE PROTEIN / acetylcholine receptor / nicotine / varenicline / MEMBRANE PROTEIN/IMMUNE SYSTEM / anti-BRIL synthetic antibody / MEMBRANE PROTEIN-IMMUNE SYSTEM complex |