Premium
Site‐specific protein labeling with amine‐containing molecules using Lactobacillus plantarum sortase
Author(s) -
Matsumoto Takuya,
Takase Ryosuke,
Tanaka Tsutomu,
Fukuda Hideki,
Kondo Akihiko
Publication year - 2012
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.201100213
Subject(s) - sortase , sortase a , biochemistry , lactobacillus plantarum , chemistry , staphylococcus aureus , residue (chemistry) , peptide , amine gas treating , bacteria , combinatorial chemistry , microbiology and biotechnology , biology , organic chemistry , lactic acid , bacterial protein , gene , genetics
Modification of proteins with small molecules is a widely used and powerful tool in biological research. Enzymatic approaches are particularly promising because substrate specificity allows for site‐specific modification. Sortase A, a transpeptidase from Staphylococcus aureus , cleaves between the T and G residues in the sequence LPXTG, and subsequently links the carboxyl group of the T residue to an amino group of N‐terminal glycine oligomers by a native peptide bond. Although Gram‐positive bacteria have several kinds of sortases, there are few reports concerning their expression and substrate specificity. Here, we demonstrate site‐specific protein modification with primary amine‐containing molecules catalyzed by Lactobacillus plantarum sortase. Enhanced green fluorescent protein (EGFP) was employed as a model protein, and an amine‐containing biotin molecule was site‐specifically conjugated with LPQTSEQ‐tagged EGFP. We developed a novel Lactobacillus plantarum sortase that has different substrate specificity compared to Staphylococcus aureus sortase. Amine‐directed protein modification was achieved using the Lactobacillus plantarum sortase ''LPQTSEQ'' sequence original recognition tag. Our results demonstrate a promising method for expanding the capabilities of site‐specific protein‐small molecule modification.