Coexpression with collagen markedly increases the half-life of the recombinant human prolyl 4-hydroxylase tetramer in the yeast Pichia pastoris.
Recent coexpression studies of the subunits of human prolyl 4-hydroxylase (4-PH) in the yeast Pichia pastoris have indicated that only a minor fraction of them were present in the alpha2beta2 tetramer, while coexpression with type III procollagen markedly increased their assembly level. We report here that the half-life of the recombinant 4-PH tetramer in Pichia when studied by pulse-chase experiments was only 50 min. Coexpression with the pro alpha1(III) chains increased this half-life to 12.5 h. Coexpression with the pro alpha1(I) chains, which were produced at half the level of the pro alpha1(III) chains, gave a half-life of 6.5 h. Coexpression with collagen thus markedly increases the half-life of the 4-PH tetramer, and the half-life may be related to the level of collagen expression.
Vuorela
A
A
Department of Medical Biochemistry, University of Oulu, Finland.
Myllyharju
J
J
Pihlajaniemi
T
T
Kivirikko
K I
KI
D020134:Catalytic Domain; D004795:Enzyme Stability; D015870:Gene Expression; D006207:Half-Life; D006801:Humans; D066298:In Vitro Techniques; D010843:Pichia / Q000235:genetics; D011347:Procollagen / Q000235:genetics* / Q000378:metabolism*; D011393:Procollagen-Proline Dioxygenase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D011347:Procollagen; D011994:Recombinant Proteins; D011393:Procollagen-Proline Dioxygenase