Ni2+ Catalyzed Cleavage of TrpLE‐Fused Small Transmembrane Peptides. (9th December 2021)

• Record Type: Journal Article
• Authors: Tang, Meng; Cao, Ruiyu; Du, Lingyu; Xu, Jikang; Wu, Bin; OuYang, Bo
• Is Part Of: Chembiochem. Volume 23:Number 2(2022)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2021-12-09
• Subjects: NMR spectroscopy -- single-pass transmembrane proteins -- SNAC-tag -- transmembrane domain -- TrpLE; Biochemistry -- Periodicals; Molecular biology -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1002/cbic.202100514
• Languages: English
• ISSNs: 1439-4227

Abstract

Abstract: In addition to a membrane anchor, the transmembrane domain (TMD) of single‐pass transmembrane proteins (SPTMPs) recently has shown essential roles in the cross‐membrane activity or receptor assembly/clustering. However, these small TMD peptides are generally hydrophobic and dynamic, difficult to be expressed and purified. Here, we have integrated the power of TrpLE fusion protein and a sequence‐specific nickel‐assisted cleavage (SNAC)‐tag to produce small TMD peptides in a highly efficient way under mild conditions, which uses Ni 2+ as the cleavage reagent, avoiding the usage of toxic cyanogen bromide (CNBr). Furthermore, this method simplifies the downstream protein purification and reconstitution. Two representative TMDs, including the Spike‐TMD from severe acute respiratory syndrome coronavirus 2 (SARS2), were successfully produced with high‐quality nuclear magnetic resonance (NMR) spectra. Therefore, our study provides a more efficient and practical approach for general structural characterization of the small TM proteins. Abstract : Sequence‐specific nickel‐assisted cleavage (SNAC) of TrpLE‐fused transmembrane (TM) peptides avoids the usage of toxic cyanogen bromide and produces small TM peptides in a highly efficient way under mild conditions.