Optimized Legionella expression strain for affinity purification of His-tagged membrane proteins eliminates major multimeric contaminant

Optimized Legionella expression strain for affinity purification of His-tagged membrane proteins eliminates major multimeric contaminant

ABSTRACT Polyhistidine tags are frequently used for isolating proteins through nickel-nitrilotriacetic acid (Ni-NTA) affinity purification. However, proteins rich in histidine can also bind to the Ni-NTA resin, leading to contamination of the purification with undesired proteins. While attempting to...

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Main Authors: Sukhithasri Vijayrajratnam, Jonasz B. Patkowski, Joshua Khorsandi, Wandy L. Beatty, Shanmugapriya Kannaiah, Ahmet Hasanovic, Tamara J. O'Connor, Tiago R. D. Costa, Joseph P. Vogel
Format: Article
Language:English
Published: American Society for Microbiology 2025-07-01
Series:Microbiology Spectrum
Subjects:
polyhistidine epitope tagged proteins
membrane proteins
Legionella pneumophila
single particle analysis
cryo-EM
protein purification
Online Access:https://journals.asm.org/doi/10.1128/spectrum.03222-24
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https://journals.asm.org/doi/10.1128/spectrum.03222-24

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