A Y-Shaped Three-Arm Structure for Probing Bivalent Interactions between Protein Receptor-Ligand Using AFM and SPR

Subhadip Senapati, Sudipta Biswas, Saikat Manna, Robert Ros, Stuart Lindsay, Peiming Zhang

Research output: Contribution to journalArticle

Abstract

The goal of this research was to develop linkage chemistry for the study of bivalent interactions between a receptor and its ligand using atomic force microscopy (AFM) and surface plasmon resonance (SPR). We conceived a three-arm structure composed of flexible chains connected to a large rigid core with orthogonal functional groups at their ends for formation and attachment (or immobilization) of bivalent ligands. To demonstrate the principle, we chose the well-known biotin-streptavidin interaction as a model system. On the basis of a crystal structure of the biotin-streptavidin complex, we designed and synthesized a bisbiotin ligand to have a Y shape with two biotin motifs on its arms for binding and a functional group on its stem for immobilization or attachment, referred to as y-bisbiotin. First, we found that the y-bisbiotin ligand stabilized the streptavidin more than its monobiotin counterpart did in solution, which indicates that the bivalent interaction was synergistic. The y-bisbiotin was attached to AFM tips through a click reaction for the force measurement experiments, which showed that unbinding the bisbiotin from streptavidin needed twice the force of unbinding a monobiotin. For the SPR study, we added a ω-thiolated alkyl chain to y-bisbiotin for its incorporation into a monolayer. The SPR data indicated that the streptavidin dissociated from a mixed monolayer bearing y-bisbiotin much slower than from the one bearing monobiotin. This work demonstrates unique chemistry for the study of bivalent interactions using AFM and SPR.

Original languageEnglish (US)
Pages (from-to)6930-6940
Number of pages11
JournalLangmuir
Volume34
Issue number23
DOIs
StatePublished - Jun 12 2018

Fingerprint

Streptavidin
Surface plasmon resonance
surface plasmon resonance
biotin
Atomic force microscopy
Bearings (structural)
Ligands
atomic force microscopy
proteins
Proteins
ligands
Biotin
immobilization
Functional groups
attachment
Monolayers
interactions
chemistry
Force measurement
stems

ASJC Scopus subject areas

  • Materials Science(all)
  • Condensed Matter Physics
  • Surfaces and Interfaces
  • Spectroscopy
  • Electrochemistry

Cite this

A Y-Shaped Three-Arm Structure for Probing Bivalent Interactions between Protein Receptor-Ligand Using AFM and SPR. / Senapati, Subhadip; Biswas, Sudipta; Manna, Saikat; Ros, Robert; Lindsay, Stuart; Zhang, Peiming.

In: Langmuir, Vol. 34, No. 23, 12.06.2018, p. 6930-6940.

Research output: Contribution to journalArticle

Senapati, Subhadip ; Biswas, Sudipta ; Manna, Saikat ; Ros, Robert ; Lindsay, Stuart ; Zhang, Peiming. / A Y-Shaped Three-Arm Structure for Probing Bivalent Interactions between Protein Receptor-Ligand Using AFM and SPR. In: Langmuir. 2018 ; Vol. 34, No. 23. pp. 6930-6940.
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