TY - JOUR
T1 - Expression of human tyrosine hydroxylase type I in Escherichia coli as a protease-cleavable fusion protein
AU - Nakashima, A.
AU - Mori, K.
AU - Nagatsu, T.
AU - Ota, A.
PY - 1999
Y1 - 1999
N2 - Wild-type and N-terminal 35-, 38-, and 44-amino acid-deleted mutants of human tyrosine hydroxylase type 1 (hTH1) fused to maltose-binding protein via the target sequence for a restriction protease were expressed in Escherichia coli and purified. The fused protein was treated with the restriction protease factor Xa or enterokinase to isolate hTH1 from the fused form. The treatment of fused wild-type and 35-amino acid-deleted mutant with factor Xa and enterokinase caused non-specific cleavages in the vicinity of the phosphorylation sites, Ser19 and Ser40, due to the flexible conformation of the N-terminus of hTH1.
AB - Wild-type and N-terminal 35-, 38-, and 44-amino acid-deleted mutants of human tyrosine hydroxylase type 1 (hTH1) fused to maltose-binding protein via the target sequence for a restriction protease were expressed in Escherichia coli and purified. The fused protein was treated with the restriction protease factor Xa or enterokinase to isolate hTH1 from the fused form. The treatment of fused wild-type and 35-amino acid-deleted mutant with factor Xa and enterokinase caused non-specific cleavages in the vicinity of the phosphorylation sites, Ser19 and Ser40, due to the flexible conformation of the N-terminus of hTH1.
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U2 - 10.1007/s007020050202
DO - 10.1007/s007020050202
M3 - Article
C2 - 10599864
AN - SCOPUS:0032749033
SN - 0300-9564
VL - 106
SP - 819
EP - 824
JO - Journal of Neural Transmission
JF - Journal of Neural Transmission
IS - 9-10
ER -
