Total chemical synthesis of the B1 domain of protein L from Peptostreptococcus magnus.
Reported here is a native chemical ligation strategy for the total chemical synthesis of the B1 domain of protein L. A synthetic construct of this 76 amino acid protein domain was prepared by the chemoselective ligation of two unprotected polypeptide fragments, one containing an N-terminal cysteine residue and one containing a C-terminal thioester moiety. The polypeptide fragments utilized in the ligation reaction were readily prepared by stepwise solid phase peptide synthesis (SPPS) methods for Boc-chemistry. The milligram quantities of protein required for conventional biophysical studies were readily accessible using the synthetic protocol described here. The folding properties of the synthetic protein L construct were also determined and found to be very similar to those of a similar wild-type protein L constructs prepared by recombinant-DNA methods. This work facilitates future unnatural amino acid mutagenesis experiments on this model protein system to further dissect the molecular basis of its folding and stability.
Yang
Xiaoye
X
Department of Chemistry, Duke University, Durham, NC 27708, USA.
Fitzgerald
Michael C
MC
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000138:chemical synthesis* / Q000737:chemistry; D007700:Kinetics; D008969:Molecular Sequence Data; D010462:Peptostreptococcus / Q000737:chemistry*; D013816:Thermodynamics; D013997:Time Factors
D001426:Bacterial Proteins; C055047:Ig L-binding protein, Peptostreptococcus