Premium
A single step purification for recombinant proteins Characterization of a microtubule associated protein (MAP 2) fragment which associates with the type II cAMP‐dependent protein kinase
Author(s) -
Stofko-Hahn Renata E.,
Carr Daniel W.,
Scott John D.
Publication year - 1992
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(92)80458-s
Subject(s) - tandem affinity purification , recombinant dna , affinity chromatography , protein kinase a , biochemistry , calmodulin , fusion protein , peptide , flag tag , microbiology and biotechnology , protein purification , biology , egta , chemistry , phosphorylation , enzyme , gene , calcium , organic chemistry
A 167 base pair DNA cassette has been constructed to facilitate the detection and purification of recombinant proteins. This cassette, kfc, encodes three distinct peptide units: a phosphorylation site for the cAMP‐depandent protein kinase (PKA), called emptide, a actor Xa cleavage site, and a almodulin‐binding peptide. Expressed kfc fusion proteins can be purified from bacterial lysates in one step by affinity chromatography on calmodulin‐agarose using EGTA as eluant. As a test of this system, we describe the expression, purification and characterization of the PKA binding domain of the microtubule associated protein (MAP 2).