Products/Services Used | Details | Operation |
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Gene Synthesis> | DNA sequences encoding model SAH domains were synthesized (GeneAr t; GenScript) and subcloned into the pET28a SUMO vector (received as a kind gift from Dr Thomas Edwards) to introduce an N-terminal His-tag and SUMO fusion protein for increased expression and solubility as described5. | Get A Quote |
Naturally-occurring single α-helices (SAHs), are rich in Arg (R), Glu (E) and Lys (K) residues, and stabilized by multiple salt bridges. Understanding how salt bridges promote their stability is challenging as SAHs are long and their sequences highly variable. Thus, we designed and tested simple de novo 98-residue polypeptides containing 7-residue repeats (AEEEXXX, where X is K or R) expected to promote salt-bridge formation between Glu and Lys/Arg. Lys-rich sequences (EK3 (AEEEKKK) and EK2R1 (AEEEKRK)) both form SAHs, of which EK2R1 is more helical and thermo-stable suggesting Arg increases stability. Substituting Lys with Arg (or vice versa) in the naturally-occurring myosin-6 SAH similarly incre... More