TY - JOUR
T1 - Insertional-fusion of basic fibroblast growth factor endowed ribonuclease 1 with enhanced cytotoxicity by steric blockade of inhibitor interaction
AU - Tada, Hiroko
AU - Onizuka, Masayuki
AU - Muraki, Kazuko
AU - Masuzawa, Wataru
AU - Futami, Junichiro
AU - Kosaka, Megumi
AU - Seno, Masaharu
AU - Yamada, Hidenori
N1 - Funding Information:
This work was partly supported by a Grant-in-Aid for Okayama Foundation for Science and Technology and a Grand-in-Aid for Scientific Research from the Ministry of Education, Science, Sports and Culture of Japan.
PY - 2004/6/18
Y1 - 2004/6/18
N2 - 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.
AB - 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.
KW - Basic fibroblast growth factor
KW - CL-RNase1, 4-118 cross-linked RNase1 mutant
KW - Insertional-fusion protein
KW - Protein engineering
KW - RFNs, insertional-fusion proteins between hRNase1 and bFGF
KW - Ribonuclease
KW - Targeting
KW - bFGF, basic fibroblast growth factor
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U2 - 10.1016/j.febslet.2004.05.007
DO - 10.1016/j.febslet.2004.05.007
M3 - Article
C2 - 15196917
AN - SCOPUS:2942598158
VL - 568
SP - 39
EP - 43
JO - FEBS Letters
JF - FEBS Letters
SN - 0014-5793
IS - 1-3
ER -