Mao Hongyuan, Hart Scott A, Schink Amy, Pollok Brian A
Ansata Therapeutics, Inc. 505 Coast Boulevard South, La Jolla, California 92037, USA.
J Am Chem Soc. 2004 Mar 10;126(9):2670-1. doi: 10.1021/ja039915e.
Sortase (SrtA), a transpeptidase from Staphylococcus aureus, catalyzes a cell-wall sorting reaction at an LPXTG motif by cleaving between threonine and glycine and subsequently joining the carboxyl group of threonine to an amino group of pentaglycine on the cell wall peptidoglycan. We have applied this transpeptidyl activity of sortase to in vitro protein ligation. We found that in the presence of sortase, protein/peptide with an LPXTG motif can be specifically ligated to an aminoglycine protein/peptide via an amide bond. Additionally, sortase can even conjugate substrates such as (d)-peptides, synthetic branched peptides, and aminoglycine-derivatized small molecules to the C terminus of a recombinant protein. The sortase-mediate protein ligation is robust, specific, and easy to perform, and can be widely applied to specific protein conjugation with polypeptides or molecules of unique biochemical and biophysical properties.
分选酶（SrtA）是一种来自金黄色葡萄球菌的转肽酶，它通过在苏氨酸和甘氨酸之间切割，催化LPXTG基序处的细胞壁分选反应，随后将苏氨酸的羧基与细胞壁肽聚糖上的五聚甘氨酸的氨基连接起来。我们已将分选酶的这种转肽基活性应用于体外蛋白质连接。我们发现，在分选酶存在的情况下，带有LPXTG基序的蛋白质/肽可以通过酰胺键特异性地连接到氨基甘氨酸蛋白质/肽上。此外，分选酶甚至可以将诸如（d）-肽、合成支链肽和氨基甘氨酸衍生的小分子等底物与重组蛋白的C末端偶联。分选酶介导的蛋白质连接稳健、特异且易于操作，可广泛应用于与具有独特生化和生物物理性质的多肽或分子进行特异性蛋白质偶联。
