A recombinant homotrimer of type I procollagen that lacks the central two D-periods. The thermal stability of the triple helix is decreased by 2 to 4 degrees C.
A D-period cassette system was developed that can be used to synthesize a variety of recombinant homotrimers of type I procollagen. A construct lacking the central two D-periods of pro alpha 1(I) chains was assembled and expressed as a recombinant protein in the mammalian cell line. The recombinant protein was purified to homogeneity and the thermal stability of the triple helix assayed by rapid protease digestion. The results indicated that deletion of the central 468 amino acids from the major triple helix lowered the thermal stability of the protein by 2 to 4 degrees C. The results therefore begin to define regions of the molecule that vary in their contributions to helical stability.
Zafarullah
K
K
Department of Biochemistry and Molecular Biology, Jefferson Institute of Molecular Medicine, Jefferson Medical College, Philadelphia, Pennsylvania, USA.
Sieron
A L
AL
Fertala
A
A
Tromp
G
G
Kuivaniemi
H
H
Prockop
D J
DJ
D055395:Bone Morphogenetic Protein 1; D019485:Bone Morphogenetic Proteins; D002460:Cell Line; D003094:Collagen / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000138:chemical synthesis / Q000235:genetics; D006801:Humans; D008666:Metalloendopeptidases / Q000378:metabolism; D010748:Phosphopeptides / Q000235:genetics / Q000378:metabolism; D011347:Procollagen / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D011348:Procollagen N-Endopeptidase / Q000378:metabolism; D017433:Protein Structure, Secondary; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis / Q000737:chemistry* / Q000378:metabolism*; D013696:Temperature
D019485:Bone Morphogenetic Proteins; C062121:N-propeptide type I collagen; D010748:Phosphopeptides; D011347:Procollagen; D011993:Recombinant Fusion Proteins; D003094:Collagen; D008666:Metalloendopeptidases; D011348:Procollagen N-Endopeptidase; C525705:BMP1 protein, human; D055395:Bone Morphogenetic Protein 1