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Insertional‐fusion of basic fibroblast growth factor endowed ribonuclease 1 with enhanced cytotoxicity by steric blockade of inhibitor interaction
Author(s) -
Tada Hiroko,
Onizuka Masayuki,
Muraki Kazuko,
Masuzawa Wataru,
Futami Junichiro,
Kosaka Megumi,
Seno Masaharu,
Yamada Hidenori
Publication year - 2004
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2004.05.007
Subject(s) - fusion protein , fusion gene , basic fibroblast growth factor , rnase p , chemistry , insertional mutagenesis , microbiology and biotechnology , ribonuclease , cell fusion , fgf10 , growth factor , biology , biochemistry , gene , gene expression , receptor , rna , recombinant dna , cell , genome
Basic fibroblast growth factor (bFGF) was inserted in the middle of human ribonuclease 1 (RNase1) sequence at an RNase inhibitor (RI)‐binding site (Gly89) by a new gene fusion technique, insertional‐fusion. The resultant insertional‐fusion protein (CL‐RFN89) was active both as bFGF and as RNase. Furthermore, it acquired an additional ability of evading RI through steric blockade of RI‐binding caused by fused bFGF domain. As a result, CL‐RFN89 showed stronger growth inhibition on B16/BL6 melanoma cells than an RI‐sensitive tandem fusion protein. Thus, the insertional‐fusion technique increases accessible positions for gene fusion on RNase, resulting in construction of a potent cytotoxic RNase.