Synthesis and conformational properties of a recombinant C-propeptide of human type III procollagen.
A cDNA was prepared that coded for the signal peptide of type III procollagen linked to the complete C-propeptide of the protein. The cDNA was then used to express the protein in a baculovirus recombinant system. Recombinant protein was recovered as a trimer from the medium of transfected cells in a yield of 1 to 2.5 mg per liter. Mapping of peptide fragments with and without reduction indicated that the protein contained the expected interchain disulfide bonds. Analysis by circular dichroism suggested that the conformation of the protein corresponded to the native conformation. Therefore, the protein should be appropriate for further tests of its biological function and analysis of structure by X-ray diffraction.
Zafarullah
K
K
Department of Biochemistry and Molecular Biology, Jefferson Institute of Molecular Medicine, Jefferson Medical College, Thomas Jefferson University, Philadelphia, USA.
Brown
E M
EM
Kuivaniemi
H
H
Tromp
G
G
Sieron
A L
AL
Fertala
A
A
Prockop
D J
DJ
D000818:Animals; D016367:Baculoviridae / Q000378:metabolism; D002478:Cells, Cultured; D002645:Chickens; D002942:Circular Dichroism; D018076:DNA, Complementary / Q000378:metabolism; D006801:Humans; D007313:Insecta / Q000378:metabolism / Q000821:virology; D011347:Procollagen / Q000096:biosynthesis* / Q000737:chemistry*; D011487:Protein Conformation; D011498:Protein Precursors / Q000096:biosynthesis* / Q000737:chemistry; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000737:chemistry; D013997:Time Factors
D018076:DNA, Complementary; D011347:Procollagen; D011498:Protein Precursors; D011994:Recombinant Proteins