Evidence for Synthesis of Alkaline Phosphatase on Membrane‐Bound Polysomes in Escherichia coli
Stanislas Varenne, Michel Piovant, Jean‐Marie Pagès, Claude J. Lazdunski
Abstract
Stanislas Varenne, Michel Piovant, Jean‐Marie Pagès, Claude J. Lazdunski
Abstract
Polysomes containing nascent chains of alkaline phosphatase have been isolated from a membrane-bound polysome preparation. Indirect immunoprecipitation using conformation-specific antibodies has been employed. This technique provides a good enrichment of these polyribosomes since routinely no more than than 10--15% of non-specific immunoprecipitation was observed. The yield of the procedure is generally 40% but can be increased if higher non-specific immunoprecipitation is tolerated. Antibodies, previously described, directed against uncoiled or folded monomers of alkaline phosphatase can be used as primary antibody to recognize the nascent chains contained in membrane-bound polysomes which suggests that these chains are partially folded.
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Polysomes containing nascent chains of alkaline phosphatase have been isolated from a membrane-bound polysome preparation. Indirect immunoprecipitation using conformation-specific antibodies has been employed. This technique provides a good enrichment of these polyribosomes since routinely no more than than 10--15% of non-specific immunoprecipitation was observed. The yield of the procedure is generally 40% but can be increased if higher non-specific immunoprecipitation is tolerated. Antibodies, previously described, directed against uncoiled or folded monomers of alkaline phosphatase can be used as primary antibody to recognize the nascent chains contained in membrane-bound polysomes which suggests that these chains are partially folded.
Key concepts: Polysome, Immunoprecipitation, Alkaline phosphatase, Phosphatase, Membrane, Biochemistry, Chemistry, Antibody