Insertional-fusion of basic fibroblast growth factor endowed ribonuclease 1 with enhanced cytotoxicity by steric blockade of inhibitor interaction

Hiroko Tada; Masayuki Onizuka; Kazuko Muraki; Wataru Masuzawa; Junichiro Futami; Megumi Kosaka; Masaharu Seno; Hidenori Yamada

doi:10.1016/j.febslet.2004.05.007

Insertional-fusion of basic fibroblast growth factor endowed ribonuclease 1 with enhanced cytotoxicity by steric blockade of inhibitor interaction

Hiroko Tada, Masayuki Onizuka, Kazuko Muraki, Wataru Masuzawa, Junichiro Futami, Megumi Kosaka, Masaharu Seno, Hidenori Yamada

Research output: Contribution to journal › Article

9 Citations (Scopus)

Abstract

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.

Original language	English
Pages (from-to)	39-43
Number of pages	5
Journal	FEBS Letters
Volume	568
Issue number	1-3
DOIs	https://doi.org/10.1016/j.febslet.2004.05.007
Publication status	Published - Jun 18 2004

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Keywords

Basic fibroblast growth factor
bFGF, basic fibroblast growth factor
CL-RNase1, 4-118 cross-linked RNase1 mutant
Insertional-fusion protein
Protein engineering
RFNs, insertional-fusion proteins between hRNase1 and bFGF
Ribonuclease
Targeting

ASJC Scopus subject areas

Biochemistry
Biophysics
Molecular Biology

Cite this

Tada, H., Onizuka, M., Muraki, K., Masuzawa, W., Futami, J., Kosaka, M., Seno, M., & Yamada, H. (2004). Insertional-fusion of basic fibroblast growth factor endowed ribonuclease 1 with enhanced cytotoxicity by steric blockade of inhibitor interaction. FEBS Letters, 568(1-3), 39-43. https://doi.org/10.1016/j.febslet.2004.05.007

Insertional-fusion of basic fibroblast growth factor endowed ribonuclease 1 with enhanced cytotoxicity by steric blockade of inhibitor interaction. / Tada, Hiroko; Onizuka, Masayuki; Muraki, Kazuko; Masuzawa, Wataru; Futami, Junichiro ; Kosaka, Megumi ; Seno, Masaharu; Yamada, Hidenori.

In: FEBS Letters, Vol. 568, No. 1-3, 18.06.2004, p. 39-43.

Research output: Contribution to journal › Article

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title = "Insertional-fusion of basic fibroblast growth factor endowed ribonuclease 1 with enhanced cytotoxicity by steric blockade of inhibitor interaction",

abstract = "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.",

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AU - Tada, Hiroko

AU - Onizuka, Masayuki

AU - Muraki, Kazuko

AU - Masuzawa, Wataru

AU - Futami, Junichiro

AU - Kosaka, Megumi

AU - Seno, Masaharu

AU - Yamada, Hidenori

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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.

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Access to Document

10.1016/j.febslet.2004.05.007