Fibronectin conformation switch induced by coadsorption with human serum albumin

Nicoletta Giamblanco, Mohammed Yaseen, Genady Zhavnerko, Jian R. Lu, Giovanni Marletta

Research output: Contribution to journalArticle

Abstract

The dynamic adsorption of human serum albumin (HSA) and plasma fibronectin (Fn) onto hydrophobic poly-(hydroxymethylsiloxane) (PHMS) and the structures of adsorbed protein layers from single and binary protein solutions were studied. Spectroscopic ellipsometry (SE) and quartz crystal microbalance with dissipation monitoring (QCM-D) together with atomic force microscopy (AFM) were used to measure the effective mass, thickness, viscoelastic properties, and morphology of the adsorbed protein films. Adsorbed HSA formed a rigid, tightly bound monolayer of deformed protein, and Fn adsorption yielded a thick, very viscoelastic layer that was firmly bound to the substrate. The mixed protein layers obtained from the coadsorption of binary equimolecular HSA-Fn solutions were found to be almost exclusively dominated by Fn molecules. Further sequential adsorption experiments showed little evidence of HSA adsorbed onto the predeposited Fn layer (denoted as Fn >> HSA), and Fn was not adsorbed onto predeposited HSA (HSA >> Fn). The conformational arrangement of the adsorbed Fn was analyzed in terms of the relative availability of two Fn domains. In particular, (4)F(1)center dot(5)F(1) binding domains in the Hep I fragment, close to the amino terminal of Fit, were targeted using a polyclonal antifibronectin antibody (anti-Fn), and the RGD sequence in the 10th segment, in the central region of the molecule, was tested by cell culture experiments. The results suggested that coadsorption with HSA induced the Fn switch from an open conformation, with the amino terminal subunit oriented toward the solution, to a close conformation, with the Fn central region oriented toward the solution.
Original languageEnglish
Pages (from-to)312-319
Number of pages8
JournalLangmuir
Volume27
Issue number1
DOIs
Publication statusPublished - 2011
Externally publishedYes

Cite this

Giamblanco, Nicoletta ; Yaseen, Mohammed ; Zhavnerko, Genady ; Lu, Jian R. ; Marletta, Giovanni. / Fibronectin conformation switch induced by coadsorption with human serum albumin. In: Langmuir. 2011 ; Vol. 27, No. 1. pp. 312-319.
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abstract = "The dynamic adsorption of human serum albumin (HSA) and plasma fibronectin (Fn) onto hydrophobic poly-(hydroxymethylsiloxane) (PHMS) and the structures of adsorbed protein layers from single and binary protein solutions were studied. Spectroscopic ellipsometry (SE) and quartz crystal microbalance with dissipation monitoring (QCM-D) together with atomic force microscopy (AFM) were used to measure the effective mass, thickness, viscoelastic properties, and morphology of the adsorbed protein films. Adsorbed HSA formed a rigid, tightly bound monolayer of deformed protein, and Fn adsorption yielded a thick, very viscoelastic layer that was firmly bound to the substrate. The mixed protein layers obtained from the coadsorption of binary equimolecular HSA-Fn solutions were found to be almost exclusively dominated by Fn molecules. Further sequential adsorption experiments showed little evidence of HSA adsorbed onto the predeposited Fn layer (denoted as Fn >> HSA), and Fn was not adsorbed onto predeposited HSA (HSA >> Fn). The conformational arrangement of the adsorbed Fn was analyzed in terms of the relative availability of two Fn domains. In particular, (4)F(1)center dot(5)F(1) binding domains in the Hep I fragment, close to the amino terminal of Fit, were targeted using a polyclonal antifibronectin antibody (anti-Fn), and the RGD sequence in the 10th segment, in the central region of the molecule, was tested by cell culture experiments. The results suggested that coadsorption with HSA induced the Fn switch from an open conformation, with the amino terminal subunit oriented toward the solution, to a close conformation, with the Fn central region oriented toward the solution.",
author = "Nicoletta Giamblanco and Mohammed Yaseen and Genady Zhavnerko and Lu, {Jian R.} and Giovanni Marletta",
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Giamblanco, N, Yaseen, M, Zhavnerko, G, Lu, JR & Marletta, G 2011, 'Fibronectin conformation switch induced by coadsorption with human serum albumin' Langmuir, vol. 27, no. 1, pp. 312-319. https://doi.org/10.1021/la104127q

Fibronectin conformation switch induced by coadsorption with human serum albumin. / Giamblanco, Nicoletta; Yaseen, Mohammed; Zhavnerko, Genady; Lu, Jian R.; Marletta, Giovanni.

In: Langmuir, Vol. 27, No. 1, 2011, p. 312-319.

Research output: Contribution to journalArticle

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T1 - Fibronectin conformation switch induced by coadsorption with human serum albumin

AU - Giamblanco, Nicoletta

AU - Yaseen, Mohammed

AU - Zhavnerko, Genady

AU - Lu, Jian R.

AU - Marletta, Giovanni

PY - 2011

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N2 - The dynamic adsorption of human serum albumin (HSA) and plasma fibronectin (Fn) onto hydrophobic poly-(hydroxymethylsiloxane) (PHMS) and the structures of adsorbed protein layers from single and binary protein solutions were studied. Spectroscopic ellipsometry (SE) and quartz crystal microbalance with dissipation monitoring (QCM-D) together with atomic force microscopy (AFM) were used to measure the effective mass, thickness, viscoelastic properties, and morphology of the adsorbed protein films. Adsorbed HSA formed a rigid, tightly bound monolayer of deformed protein, and Fn adsorption yielded a thick, very viscoelastic layer that was firmly bound to the substrate. The mixed protein layers obtained from the coadsorption of binary equimolecular HSA-Fn solutions were found to be almost exclusively dominated by Fn molecules. Further sequential adsorption experiments showed little evidence of HSA adsorbed onto the predeposited Fn layer (denoted as Fn >> HSA), and Fn was not adsorbed onto predeposited HSA (HSA >> Fn). The conformational arrangement of the adsorbed Fn was analyzed in terms of the relative availability of two Fn domains. In particular, (4)F(1)center dot(5)F(1) binding domains in the Hep I fragment, close to the amino terminal of Fit, were targeted using a polyclonal antifibronectin antibody (anti-Fn), and the RGD sequence in the 10th segment, in the central region of the molecule, was tested by cell culture experiments. The results suggested that coadsorption with HSA induced the Fn switch from an open conformation, with the amino terminal subunit oriented toward the solution, to a close conformation, with the Fn central region oriented toward the solution.

AB - The dynamic adsorption of human serum albumin (HSA) and plasma fibronectin (Fn) onto hydrophobic poly-(hydroxymethylsiloxane) (PHMS) and the structures of adsorbed protein layers from single and binary protein solutions were studied. Spectroscopic ellipsometry (SE) and quartz crystal microbalance with dissipation monitoring (QCM-D) together with atomic force microscopy (AFM) were used to measure the effective mass, thickness, viscoelastic properties, and morphology of the adsorbed protein films. Adsorbed HSA formed a rigid, tightly bound monolayer of deformed protein, and Fn adsorption yielded a thick, very viscoelastic layer that was firmly bound to the substrate. The mixed protein layers obtained from the coadsorption of binary equimolecular HSA-Fn solutions were found to be almost exclusively dominated by Fn molecules. Further sequential adsorption experiments showed little evidence of HSA adsorbed onto the predeposited Fn layer (denoted as Fn >> HSA), and Fn was not adsorbed onto predeposited HSA (HSA >> Fn). The conformational arrangement of the adsorbed Fn was analyzed in terms of the relative availability of two Fn domains. In particular, (4)F(1)center dot(5)F(1) binding domains in the Hep I fragment, close to the amino terminal of Fit, were targeted using a polyclonal antifibronectin antibody (anti-Fn), and the RGD sequence in the 10th segment, in the central region of the molecule, was tested by cell culture experiments. The results suggested that coadsorption with HSA induced the Fn switch from an open conformation, with the amino terminal subunit oriented toward the solution, to a close conformation, with the Fn central region oriented toward the solution.

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JO - Langmuir

JF - Langmuir

SN - 0743-7463

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