TY - JOUR
T1 - In vitro study of proteolytic degradation of rat histidine decarboxylase
AU - Olmo, María T.
AU - Urdiales, José L.
AU - Pegg, Anthony E.
AU - Medina Torres, Miguel A.
AU - Sánchez-Jiménez, Francisca
PY - 2000
Y1 - 2000
N2 - Mammalian ornithine decarboxylase (ODC) is a very unstable protein which is degraded in an ATP-dependent manner by proteasome 26S, after making contact with the regulatory protein antizyme. PEST regions are sequences described as signals for protein degradation. The C-terminal PEST region of mammalian ODC is essential for its degradation by proteasome 26S. Mammalian histidine decarboxylase (HDC) is also a short-lived protein. The full primary sequence of mammalian HDC contains PEST-regions at both the N- and C-termini. Rat ODC and different truncated and full versions of rat HDC were expressed in vitro. In vitro degradation of rat ODC and rat 1-512 HDC were compared. Like ODC, rat 1-512 HDC is degraded mainly by an ATP-dependent mechanism. However, antizyme has no effect on the degradation of 1-512 HDC. The use of the inhibitors MG-132 and lactacystine significantly inhibited the degradation of 1-512 HDC, suggesting that a ubiquitin-dependent, proteasome 26S proteolytic pathway is involved. Results obtained with the different modifications of rat HDC containing all three PEST regions (full version, 1- 656 HDC), only the N-terminal PEST region (1-512 HDC), or no PEST region (69- 512 HDC), indicate that the N-terminal (1-69) fragment, but not the C- terminal fragment, determines that the HDC protein is a proteasome substrate in vitro.
AB - Mammalian ornithine decarboxylase (ODC) is a very unstable protein which is degraded in an ATP-dependent manner by proteasome 26S, after making contact with the regulatory protein antizyme. PEST regions are sequences described as signals for protein degradation. The C-terminal PEST region of mammalian ODC is essential for its degradation by proteasome 26S. Mammalian histidine decarboxylase (HDC) is also a short-lived protein. The full primary sequence of mammalian HDC contains PEST-regions at both the N- and C-termini. Rat ODC and different truncated and full versions of rat HDC were expressed in vitro. In vitro degradation of rat ODC and rat 1-512 HDC were compared. Like ODC, rat 1-512 HDC is degraded mainly by an ATP-dependent mechanism. However, antizyme has no effect on the degradation of 1-512 HDC. The use of the inhibitors MG-132 and lactacystine significantly inhibited the degradation of 1-512 HDC, suggesting that a ubiquitin-dependent, proteasome 26S proteolytic pathway is involved. Results obtained with the different modifications of rat HDC containing all three PEST regions (full version, 1- 656 HDC), only the N-terminal PEST region (1-512 HDC), or no PEST region (69- 512 HDC), indicate that the N-terminal (1-69) fragment, but not the C- terminal fragment, determines that the HDC protein is a proteasome substrate in vitro.
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U2 - 10.1046/j.1432-1327.2000.01153.x
DO - 10.1046/j.1432-1327.2000.01153.x
M3 - Article
C2 - 10691992
AN - SCOPUS:0034091061
SN - 0014-2956
VL - 267
SP - 1527
EP - 1531
JO - European Journal of Biochemistry
JF - European Journal of Biochemistry
IS - 5
ER -