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In Vitro PMP Import Analysis Using Cell-Free Synthesized PMP and Isolated Peroxisomes
Yuqiong Liu1, Masanori Honsho2, Yukio Fujiki3
1Graduate School of Systems Life Sciences, Kyushu University Graduate School, Nishi-ku, Fukuoka, 819-0395, Japan.
Methods in Molecular Biology (Clifton, N.J.)
|April 15, 2017
Abstract:
The Pex19p- and Pex3p-dependent direct import of peroxisomal membrane proteins (PMPs), termed the class I pathway, can be reconstituted in vitro by incubating cell-free synthesized PMPs with highly purified peroxisomes at 26 °C for 1 h. This method ensures that the proteins targeted to peroxisomes are imported directly without involvement of other organelles.
