Sortase-mediated protein ligation: A new method for protein engineering
Sortase-mediated protein ligation: A new method for protein engineering
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DOI：
10.1021/ja039915e
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发表时间：
2004-03-10
影响因子：
15
通讯作者：
Pollok, BA
中科院分区：
文献类型：
--
作者：
Mao, HY;Hart, SA;Pollok, BA
Sortase （SrtA）是一种来自金黄色葡萄球菌的转肽酶，通过在苏氨酸和甘氨酸之间切割，随后将苏氨酸的羧基与细胞壁肽聚糖上的五甘氨酸的氨基结合，在LPXTG基序上催化细胞壁分选反应。我们已经将这种排序酶的转肽基活性应用于体外蛋白连接。我们发现，在排序酶存在的情况下，具有LPXTG基序的蛋白/肽可以通过酰胺键特异性地连接到氨基甘氨酸蛋白/肽上。此外，分选酶甚至可以将底物如(d)肽、合成支链肽和氨基甘氨酸衍生的小分子偶联到重组蛋白的C端。sortase介导的蛋白质连接具有强大、特异性和易于进行的特点，可广泛应用于特定蛋白质与多肽或具有独特生化和生物物理性质的分子的结合。
Sortase (SrtA), a transpeptidase fromStaphylococcus 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.