Deletion of N-terminus of human tyrosine hydroxylase type 1 enhances stability of the enzyme in AtT-20 cells.

作者: Akira Nakashima , Nobuhiro Hayashi , Yoko S. Kaneko , Keiji Mori , Hiromi Egusa

DOI: 10.1002/JNR.20540

关键词:

摘要: Wildtype human tyrosine hydroxylase (TH) type 1 and 4 mutants (del-52, a form with the first 52 amino acid residues deleted; del-157, one 157 RR-EE, in which Arg 3 7 -Arg 8 was replaced by Glu -Glu ; S40D, Ser 0 Asp ) were expressed AtT-20 mouse neuroendocrine cells order to clarify how deeply N-terminus of TH is involved efficient production dopamine (DA) mammalian cells. The amounts DA that accumulated expressing these (hTH1) phenotypes following order: del-52 = del-157 RR-EE > S40D wildtype, although enzyme activities lower than those S40D. observation on immunoblot analyses N-terminus-deleted hTH1 much more stable wildtype can reconcile discrepant results. Computer-assisted analysis spatial configuration identified five newly recognized PEST motifs, located sequence Met -Lys 2 predicted deletion region would alter secondary structure within catalytic domain. Collectively, high stability be generated loss motif their N-termini structural change domain, promise an deleted hTH1.

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