Expression and Characterization of Phosphorylated Recombinant Human β-Casein inEscherichia coli

Specific serine and threonine residues of recombinant human β-casein produced inEscherichia coliwere shown to be phosphorylatedin vivowhen human casein kinase II was coexpressed in the same plasmid. All of the phosphorylated forms found in the native protein were also detected in the recombinant pro...

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Published in:Protein expression and purification 1997-07, Vol.10 (2), p.202-208
Main Authors: Thurmond, Jennifer M., Hards, Robert G., Seipelt, C.Thomas, Leonard, Amanda E., Hansson, Lennart, Strömqvist, Mats, Byström, Mona, Enquist, Kerstin, Xu, Bixiong C., Kopchick, John J., Mukerji, Pradip
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cited_by cdi_FETCH-LOGICAL-c1337-13e21932e259a5b321c738a73fadf625a0cd7f39a58de0837284ba24c924212b3
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container_end_page 208
container_issue 2
container_start_page 202
container_title Protein expression and purification
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creator Thurmond, Jennifer M.
Hards, Robert G.
Seipelt, C.Thomas
Leonard, Amanda E.
Hansson, Lennart
Strömqvist, Mats
Byström, Mona
Enquist, Kerstin
Xu, Bixiong C.
Kopchick, John J.
Mukerji, Pradip
description Specific serine and threonine residues of recombinant human β-casein produced inEscherichia coliwere shown to be phosphorylatedin vivowhen human casein kinase II was coexpressed in the same plasmid. All of the phosphorylated forms found in the native protein were also detected in the recombinant protein. The phosphorylation of recombinant human β-casein was confirmed by immunoblots, fast protein liquid chromatography, urea–polyacrylamide gel electrophoresis, SDS–polyacrylamide gel electrophoresis, and liquid chromatography–mass spectrometry. The results indicate that the substrate specificity of casein kinase IIin vivowas unaffected in its recombinant form. This is the first demonstration ofin vivophosphorylation of specific residues of a multiphosphorylated protein produced inE. coliwith a single plasmid.
doi_str_mv 10.1006/prep.1997.0737
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title Expression and Characterization of Phosphorylated Recombinant Human β-Casein inEscherichia coli
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