New yeast production method sharpens NMR spectra of membrane proteins at low cost

Solid-state NMR is a powerful tool for studying membrane protein structures, but high spectral resolution is often hindered by ¹³C coupling effects. To that end, researchers from East China University of Science and Technology and Peking University have now introduced a simple and cost-effective labeling method that overcomes this challenge.

For the study, the team used Pichia pastoris (P. pastoris), a yeast that can grow on methanol as its only carbon source. “By mixing natural abundance methanol with carbon-13-labeled methanol in the culture medium, we achieved random fractional labeling of a eukaryotic membrane protein—a rhodopsin from Leptosphaeria maculans (LR). This approach reduces carbon-carbon coupling, sharpening spectral peaks and improving resolution,” shares Shenlin Wang, the study’s corresponding author and a professor at East China University of Science and Technology.

“What's exciting is that we can now obtain high-resolution NMR spectra using only inexpensive ¹³C methanol,” adds Wang. “This method is particularly useful for researchers who need to assess protein quality or study site-specific dynamics without the high costs of specialized labeled precursors.”

Notably, the researchers The found that a 25% ¹³C enrichment level offered the best balance between resolution and signal sensitivity, cutting average ¹³C linewidths by half compared to fully labeled proteins. This led to a 50% increase in well-resolved peaks in key NMR spectra. Importantly, the spectral quality matched that achieved with far more expensive 2-¹³C glycerol labeling—but at about one-tenth of the cost.

“This labeling strategy is expected to facilitate structural, dynamic, and interaction studies of eukaryotic membrane proteins, which are crucial for understanding many biological processes and drug targets,” Wang says.
 

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Contact the author: Shenlin Wang, East China University of Science and Technology, wangshenlin@ecust.edu.cn

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