The heparin binding motif of endostatin mediates its interaction with receptor nucleolin

Yan Fu, Yang Chen, Xu Luo, Yun Liang, Hubing Shi, Lei Gao, Shunli Zhan, Daifu Zhou, Yongzhang Luo

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Endostatin is a potent angiogenesis inhibitor with heparin-dependent activities. Nucleolin, a novel functional receptor of endostatin, mediates both the internalization to endothelial cells and the antiangiogenic activity of endostatin. To define the exact role of the heparin binding motif in mediating the interaction between endostatin and its receptor nucleolin, up to six arginine residues (R155, R158, R184, R270, R193, and R194) located in the heparin binding motif of endostatin were substituted by alanine to make double, quadruple, or hexad point mutations, respectively. Contributions of the heparin binding motif to both the interaction with nucleolin and the biological activities of endostatin were investigated from in vitro to in vivo. Here we show that Arg to Ala point mutagenesis of the heparin binding motif does not interrupt the folding of endostatin but significantly impairs the interaction between endostatin and nucleolin. Double and quadruple mutants showed significantly decreased internalization to endothelial cells and antitumor activities, while the hexad Arg to Ala mutant completely lost its interaction with nucleolin and biological functions. Taken together, the present study demonstrates that the arginine clusters in the heparin binding motif of endostatin significantly contribute to its interaction with receptor nucleolin and mediate the antiangiogenic and antitumor activities of endostatin.

Original languageEnglish (US)
Pages (from-to)11655-11663
Number of pages9
JournalBiochemistry
Volume48
Issue number49
DOIs
StatePublished - Dec 15 2009

Fingerprint

Endostatins
Heparin
Endothelial cells
Arginine
Endothelial Cells
nucleolin
Mutagenesis
Angiogenesis Inhibitors
Bioactivity
Point Mutation
Alanine

ASJC Scopus subject areas

  • Biochemistry

Cite this

The heparin binding motif of endostatin mediates its interaction with receptor nucleolin. / Fu, Yan; Chen, Yang; Luo, Xu; Liang, Yun; Shi, Hubing; Gao, Lei; Zhan, Shunli; Zhou, Daifu; Luo, Yongzhang.

In: Biochemistry, Vol. 48, No. 49, 15.12.2009, p. 11655-11663.

Research output: Contribution to journalArticle

Fu, Y, Chen, Y, Luo, X, Liang, Y, Shi, H, Gao, L, Zhan, S, Zhou, D & Luo, Y 2009, 'The heparin binding motif of endostatin mediates its interaction with receptor nucleolin', Biochemistry, vol. 48, no. 49, pp. 11655-11663. https://doi.org/10.1021/bi901265z
Fu, Yan ; Chen, Yang ; Luo, Xu ; Liang, Yun ; Shi, Hubing ; Gao, Lei ; Zhan, Shunli ; Zhou, Daifu ; Luo, Yongzhang. / The heparin binding motif of endostatin mediates its interaction with receptor nucleolin. In: Biochemistry. 2009 ; Vol. 48, No. 49. pp. 11655-11663.
@article{86e38e01c1f849a9a8a34b0e4f184d8e,
title = "The heparin binding motif of endostatin mediates its interaction with receptor nucleolin",
abstract = "Endostatin is a potent angiogenesis inhibitor with heparin-dependent activities. Nucleolin, a novel functional receptor of endostatin, mediates both the internalization to endothelial cells and the antiangiogenic activity of endostatin. To define the exact role of the heparin binding motif in mediating the interaction between endostatin and its receptor nucleolin, up to six arginine residues (R155, R158, R184, R270, R193, and R194) located in the heparin binding motif of endostatin were substituted by alanine to make double, quadruple, or hexad point mutations, respectively. Contributions of the heparin binding motif to both the interaction with nucleolin and the biological activities of endostatin were investigated from in vitro to in vivo. Here we show that Arg to Ala point mutagenesis of the heparin binding motif does not interrupt the folding of endostatin but significantly impairs the interaction between endostatin and nucleolin. Double and quadruple mutants showed significantly decreased internalization to endothelial cells and antitumor activities, while the hexad Arg to Ala mutant completely lost its interaction with nucleolin and biological functions. Taken together, the present study demonstrates that the arginine clusters in the heparin binding motif of endostatin significantly contribute to its interaction with receptor nucleolin and mediate the antiangiogenic and antitumor activities of endostatin.",
author = "Yan Fu and Yang Chen and Xu Luo and Yun Liang and Hubing Shi and Lei Gao and Shunli Zhan and Daifu Zhou and Yongzhang Luo",
year = "2009",
month = "12",
day = "15",
doi = "10.1021/bi901265z",
language = "English (US)",
volume = "48",
pages = "11655--11663",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "49",

}

TY - JOUR

T1 - The heparin binding motif of endostatin mediates its interaction with receptor nucleolin

AU - Fu, Yan

AU - Chen, Yang

AU - Luo, Xu

AU - Liang, Yun

AU - Shi, Hubing

AU - Gao, Lei

AU - Zhan, Shunli

AU - Zhou, Daifu

AU - Luo, Yongzhang

PY - 2009/12/15

Y1 - 2009/12/15

N2 - Endostatin is a potent angiogenesis inhibitor with heparin-dependent activities. Nucleolin, a novel functional receptor of endostatin, mediates both the internalization to endothelial cells and the antiangiogenic activity of endostatin. To define the exact role of the heparin binding motif in mediating the interaction between endostatin and its receptor nucleolin, up to six arginine residues (R155, R158, R184, R270, R193, and R194) located in the heparin binding motif of endostatin were substituted by alanine to make double, quadruple, or hexad point mutations, respectively. Contributions of the heparin binding motif to both the interaction with nucleolin and the biological activities of endostatin were investigated from in vitro to in vivo. Here we show that Arg to Ala point mutagenesis of the heparin binding motif does not interrupt the folding of endostatin but significantly impairs the interaction between endostatin and nucleolin. Double and quadruple mutants showed significantly decreased internalization to endothelial cells and antitumor activities, while the hexad Arg to Ala mutant completely lost its interaction with nucleolin and biological functions. Taken together, the present study demonstrates that the arginine clusters in the heparin binding motif of endostatin significantly contribute to its interaction with receptor nucleolin and mediate the antiangiogenic and antitumor activities of endostatin.

AB - Endostatin is a potent angiogenesis inhibitor with heparin-dependent activities. Nucleolin, a novel functional receptor of endostatin, mediates both the internalization to endothelial cells and the antiangiogenic activity of endostatin. To define the exact role of the heparin binding motif in mediating the interaction between endostatin and its receptor nucleolin, up to six arginine residues (R155, R158, R184, R270, R193, and R194) located in the heparin binding motif of endostatin were substituted by alanine to make double, quadruple, or hexad point mutations, respectively. Contributions of the heparin binding motif to both the interaction with nucleolin and the biological activities of endostatin were investigated from in vitro to in vivo. Here we show that Arg to Ala point mutagenesis of the heparin binding motif does not interrupt the folding of endostatin but significantly impairs the interaction between endostatin and nucleolin. Double and quadruple mutants showed significantly decreased internalization to endothelial cells and antitumor activities, while the hexad Arg to Ala mutant completely lost its interaction with nucleolin and biological functions. Taken together, the present study demonstrates that the arginine clusters in the heparin binding motif of endostatin significantly contribute to its interaction with receptor nucleolin and mediate the antiangiogenic and antitumor activities of endostatin.

UR - http://www.scopus.com/inward/record.url?scp=71549142577&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=71549142577&partnerID=8YFLogxK

U2 - 10.1021/bi901265z

DO - 10.1021/bi901265z

M3 - Article

VL - 48

SP - 11655

EP - 11663

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 49

ER -