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Cell‐free synthesis of alkaline lipase, a glyoxysomal membrane protein, from castor bean endosperm
Author(s) -
Maeshima Masayoshi,
Takeuchi Atsuko,
Asahi Tadashi
Publication year - 1987
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(87)80868-2
Subject(s) - endosperm , lipase , biochemistry , polysome , biology , enzyme , cell free system , membrane protein , cell free protein synthesis , protein biosynthesis , membrane , rna , gene , ribosome
Polyadenylated RNA from castor bean endosperm was translated in a wheat germ cell‐free protein synthesis system, and alkaline lipase, an integral glyoxysomal membrane protein, was immunoprecipitated. The apparent molecular mass of the lipase synthesized in vitro was slightly higher than that of the mature enzyme (62 kDa). When mRNA derived from free and membrane‐bound polysomes in the endosperms was translated in vitro, the lipase was predominantly recovered from the products of the free polysomal mRNA, suggesting that the membrane protein is post‐translationally inserted into the membrane. The amino‐terminal 8 amino acid residues of the mature lipase were sequenced.