A CELL-FREE PROTEIN TRANSLOCATION SYSTEM PREPARED ENTIRELY FROM A GRAM-POSITIVE ORGANISM

作者：SCHIMZ KL^*; DECKER G; FRINGS E; MEENS J; KLEIN M; MULLER M

来源：FEBS Letters, 1995, 362(1): 29-33.

DOI：10.1016/0014-5793(95)00180-H

• PROTEIN SECRETION
• GRAM-POSITIVE BACTERIA
• CELL-FREE TRANSLOCATION SYSTEM
• SECA
• LIPASE
• STAPHYLOCOCCUS CARNOSUS

A cell-free protein translocation system derived exclusively from a Gram-positive bacterium is described here for the first time. Highly efficient in vitro synthesis of plasmid encoded preprolipase of Staphylococcus hyicus is accomplished by coupled transcription/translation using either a cytosolic extract of S. carnosus alone or in combination with T7-RNA-polymerase. Addition of inside-out cytoplasmic membrane vesicles of S. carnosus leads to the partial conversion (processing) of preprolipase to prolipase. In addition, as shown in a protease protection assay, a significant part of preprolipase plus prolipase is translocated in vitro into the lumen of the vesicles. Translocation of preprolipase into the membrane vesicles requires the proton-motive force and the S. carnosus SecA protein.

• 出版日期1995-3-27