Hydroxylated human homotrimeric collagen I in Agrobacterium tumefaciens-mediated transient expression and in transgenic tobacco plant.
Potential contamination of animal-derived collagen with pathogens has led to the demand for safe recombinant sources of this complex molecule. In continuation of our previous work [Ruggiero et al. (2000) FEBS Lett. 469, 132-136], here we show that it is possible to produce recombinant hydroxylated homotrimeric collagen in tobacco plants that are co-transformed with a human type I collagen and a chimeric proline-4-hydroxylase (P4H). This is to our knowledge the first time that transient expression in tobacco was used to improve the quality of a recombinant protein produced in plants through co-expression with an animal cell-derived modifying enzyme. We demonstrated the functionality of the new chimeric P4H and thus improved the thermal stability of recombinant collagen I from plants to 37 degrees C.
Merle
C
C
Meristem Therapeutics, 8 rue des Frères Lumière, 63100, Clermont-Ferrand, France.
Perret
S
S
Lacour
T
T
Jonval
V
V
Hudaverdian
S
S
Garrone
R
R
Ruggiero
F
F
Theisen
M
M
D016960:Agrobacterium tumefaciens / Q000378:metabolism*; D000596:Amino Acids / Q000032:analysis; D001681:Biological Assay; D002851:Chromatography, High Pressure Liquid; D002942:Circular Dichroism; D024042:Collagen Type I / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D015870:Gene Expression; D006801:Humans; D006900:Hydroxylation; D010434:Pepsin A / Q000737:chemistry; D030821:Plants, Genetically Modified; D011393:Procollagen-Proline Dioxygenase / Q000235:genetics / Q000378:metabolism; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013696:Temperature; D014026:Nicotiana / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D014170:Transformation, Genetic
D000596:Amino Acids; D024042:Collagen Type I; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D011393:Procollagen-Proline Dioxygenase; D010434:Pepsin A