Intervesicle Cross-Linking with Integrin IIb
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- 作者：Bin Hu ; Dirk Finsinger ; Kai Peter ; Zeno Guttenberg ; Michael Bä ; rmann ; Horst Kessler ; Achim Escherich ; Luis Moroder ; Jochen Bö ; hm ; Wolfgang Baumeister ; Sen-fang Sui ; and Erich Sackmann
- 刊名：Biochemistry
- 出版年：2000
- 出版时间：October 10, 2000
- 年：2000
- 卷：39
- 期：40
- 页码：12284 - 12294
- 全文大小：329K
- 年卷期：v.39,no.40(October 10, 2000)
- ISSN：1520-4995
文摘
We report the synthesis of a new integrin 
IIb
3-specific cyclic hexapeptide that contains anArg-Gly-Asp (RGD) sequence and is coupled to a dimyristoylthioglyceryl anchor. We demonstrate thatthis ligand is useful to study specific integrin binding to membrane surfaces. With the help of biotinylatedanalogues of the peptide, a spacer of optimal length between the peptide and lipid moieties was searchedfor by evaluating the binding strength with an enzyme-coupled immunosorbent assay (ELISA) and bysurface plasmon resonance (SPR). It was found to be strongly dependent on the length of the spacerintroduced between the biotin and peptide moieties of the ligands, which consisted either of 
-aminohexanoicacid (Ahx) or of Ahx with two additional glycine units. Best results were obtained with c[Arg-Gly-Asp-D-Phe-Lys(Biot-Ahx-Gly-Gly)-Gly-] with dissociation constants of KD = 0.158 
M from ELISAand KD = 1.1 M from SPR measurements. The analogous lipopeptide, c[Arg-Gly-Asp-D-Phe-Lys([dimyristoyl-3-thioglyceryl-succinimido-propanoyl]Ahx-Gly-Gly)-Gly], was used as a membrane-anchoredintegrin ligand. It is shown by fluorescence microscopy and cryo electron microscopy that integrinreconstituted into phospholipid vesicles binds to vesicles decorated with the lipopeptide, forming regularlyspaced bridges between the two kinds of vesicles. The novel integrin-specific ligand allows establishmentof new model systems for systematic studies of the self-organization of integrin clusters and focal adhesioncomplexes.
IIb3-specific cyclic hexapeptide that contains anArg-Gly-Asp (RGD) sequence and is coupled to a dimyristoylthioglyceryl anchor. We demonstrate thatthis ligand is useful to study specific integrin binding to membrane surfaces. With the help of biotinylatedanalogues of the peptide, a spacer of optimal length between the peptide and lipid moieties was searchedfor by evaluating the binding strength with an enzyme-coupled immunosorbent assay (ELISA) and bysurface plasmon resonance (SPR). It was found to be strongly dependent on the length of the spacerintroduced between the biotin and peptide moieties of the ligands, which consisted either of -aminohexanoicacid (Ahx) or of Ahx with two additional glycine units. Best results were obtained with c[Arg-Gly-Asp-D-Phe-Lys(Biot-Ahx-Gly-Gly)-Gly-] with dissociation constants of KD = 0.158 M from ELISAand KD = 1.1 M from SPR measurements. The analogous lipopeptide, c[Arg-Gly-Asp-D-Phe-Lys([dimyristoyl-3-thioglyceryl-succinimido-propanoyl]Ahx-Gly-Gly)-Gly], was used as a membrane-anchoredintegrin ligand. It is shown by fluorescence microscopy and cryo electron microscopy that integrinreconstituted into phospholipid vesicles binds to vesicles decorated with the lipopeptide, forming regularlyspaced bridges between the two kinds of vesicles. The novel integrin-specific ligand allows establishmentof new model systems for systematic studies of the self-organization of integrin clusters and focal adhesioncomplexes.