Measuring adsorption of a hydrophobic probe with a surface plasmon resonance sensor to monitor conformational changes in immobilized proteins

Satoshi Yamaguchi, Teruhisa Mannen, Tamotsu Zako, Noriho Kamiya, Teruyuki Nagamune

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Conformational changes of proteins immobilized on solid matrices were observed by measuring the adsorption of Triton X-100 (TX), a nonionic detergent, as a hydrophobic probe with BIACORE, a biosensor that utilizes the phenomenon of surface plasmon resonance (SPR). Two kinds of proteins, α-glucosidase and lysozyme, were covalently attached to dextran matrices on the sensor surface in the flow cell and then exposed to various concentrations of TX solution. We measured SPR signal changes derived from adsorption of TX to the immobilized proteins and calculated the monolayer adsorption capacity using the Brunauer-Emmett-Teller (BET) equation. The results demonstrated that monolayer adsorption capacity is proportional to the amount of immobilized proteins. Further, the unfolding process of immobilized proteins on the sensor surface induced by guanidine hydrochloride was investigated by monitoring SPR signal increases due to the adsorption of TX to the exposed hydrophobic region of the protein. Results strongly suggested that the increase in the SPR signal reflected the formation of the agglutinative unfolded state. We expect our measuring method using the SPR sensor and TX adsorption will be a novel tool to provide conformational information regarding various proteins on solid matrices.

Original languageEnglish
Pages (from-to)1348-1354
Number of pages7
JournalBiotechnology Progress
Volume19
Issue number4
DOIs
Publication statusPublished - Jul 1 2003
Externally publishedYes

Fingerprint

Immobilized Proteins
surface plasmon resonance
Surface Plasmon Resonance
probes (equipment)
Octoxynol
Adsorption
sensors (equipment)
adsorption
monitoring
proteins
Glucosidases
Proteins
Guanidine
guanidines
Biosensing Techniques
glucosidases
Muramidase
Dextrans
biosensors
Detergents

All Science Journal Classification (ASJC) codes

  • Biotechnology

Cite this

Measuring adsorption of a hydrophobic probe with a surface plasmon resonance sensor to monitor conformational changes in immobilized proteins. / Yamaguchi, Satoshi; Mannen, Teruhisa; Zako, Tamotsu; Kamiya, Noriho; Nagamune, Teruyuki.

In: Biotechnology Progress, Vol. 19, No. 4, 01.07.2003, p. 1348-1354.

Research output: Contribution to journalArticle

Yamaguchi, Satoshi ; Mannen, Teruhisa ; Zako, Tamotsu ; Kamiya, Noriho ; Nagamune, Teruyuki. / Measuring adsorption of a hydrophobic probe with a surface plasmon resonance sensor to monitor conformational changes in immobilized proteins. In: Biotechnology Progress. 2003 ; Vol. 19, No. 4. pp. 1348-1354.
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