Ni2+ Catalyzed Cleavage of TrpLE-Fused Small Transmembrane Peptides.
Chembiochem
; 23(2): e202100514, 2022 01 19.
Article
in English
| MEDLINE | ID: covidwho-1653182
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 Ni2+ 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.
Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
Peptides
/
Recombinant Fusion Proteins
/
Nickel
Type of study:
Randomized controlled trials
Limits:
Humans
Language:
English
Journal:
Chembiochem
Journal subject:
Biochemistry
Year:
2022
Document Type:
Article
Affiliation country:
Cbic.202100514
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