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 › peer-review

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

Keywords

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
bFGF, basic fibroblast growth factor

ASJC Scopus subject areas

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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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 › peer-review

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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.",

keywords = "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, bFGF, basic fibroblast growth factor",

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

note = "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.",

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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

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.

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KW - Insertional-fusion protein

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KW - Ribonuclease

KW - Targeting

KW - bFGF, basic fibroblast growth factor

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