Structural and functional characterisation of recombinant human erythropoietin analogues.
Deletion mutants of a synthetic human erythropoietin-cDNA were transiently expressed in COS-7 cells and products analyzed using Western blots and a cell proliferation assay. Only two mutants with deletions affecting the N-terminal portion and the amino acid sequence 115-121 displayed biological activity. The exchange of hydrophilic, charged amino acids by alanine in two potential alpha-helical regions, the internal amino acid sequence 102-106 and the C-terminal sequence 154-159, causes a 2-11-fold loss of activity. The results suggest that both regions are involved in either maintaining the active structure of the hormone or interacting with the receptor.
Bittorf
T
T
Institut für Medizinische Biochemie der Medizinischen Fakultät der Universität Rostock, Germany.
Jaster
R
R
Brock
J
J
D000595:Amino Acid Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004921:Erythropoietin / Q000737:chemistry / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D017384:Sequence Deletion; D013329:Structure-Activity Relationship
D018076:DNA, Complementary; D011994:Recombinant Proteins; D004921:Erythropoietin