Kinetic analysis of his-tagged protein binding to nickel-chelating nanolipoprotein particles

Craig D. Blanchette, Nicholas O Fischer, Michele Corzett, Graham Bench, Paul D. Hoeprich

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Nanolipoprotein particles (NLPs) are discoidal self-assembling membrane mimetics that have been primarily used as a platform for the solubilization and stabilization of membrane proteins. Nickel-chelating nanolipoprotein particles (NiNLPs) containing nickel-chelating lipids (Ni-lipid) for the targeted immobilization of His-tagged proteins hold promise as carriers of hydrophilic biological molecules for a range of applications. The effect of protein loading (i.e., the number of proteins bound per NiNLP) and Ni-lipid content on the time scales and kinetics of binding are important to various applications such as vaccine development, diagnostic imaging, and drug delivery. We have immobilized hexa-His-tagged LsrB, a Yersinia pestis transport protein, onto NiNLPs to examine the effect of protein binding stoichiometry and Ni-lipid content on the time scales and kinetics of protein binding by surface plasmon resonance (SPR). Data indicate that the dissociation half-time increases with Ni-lipid content up to a molar concentration of 35% and decreases as the number of bound protein per NiNLP increases. These findings indicate that the kinetics of protein binding are highly dependent on both the number of bound protein per NiNLP and Ni-lipid content.

Original languageEnglish (US)
Pages (from-to)1321-1330
Number of pages10
JournalBioconjugate Chemistry
Volume21
Issue number7
DOIs
StatePublished - Jul 21 2010
Externally publishedYes

Fingerprint

Chelation
Nickel
Protein Binding
Kinetics
Lipids
Proteins
Membranes
Yersinia pestis
Vaccines
Surface Plasmon Resonance
Surface plasmon resonance
Diagnostic Imaging
Drug delivery
Stoichiometry
Immobilization
Carrier Proteins
Membrane Proteins
Stabilization
Imaging techniques
Molecules

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Organic Chemistry
  • Pharmaceutical Science
  • Biomedical Engineering
  • Pharmacology

Cite this

Kinetic analysis of his-tagged protein binding to nickel-chelating nanolipoprotein particles. / Blanchette, Craig D.; Fischer, Nicholas O; Corzett, Michele; Bench, Graham; Hoeprich, Paul D.

In: Bioconjugate Chemistry, Vol. 21, No. 7, 21.07.2010, p. 1321-1330.

Research output: Contribution to journalArticle

Blanchette, Craig D. ; Fischer, Nicholas O ; Corzett, Michele ; Bench, Graham ; Hoeprich, Paul D. / Kinetic analysis of his-tagged protein binding to nickel-chelating nanolipoprotein particles. In: Bioconjugate Chemistry. 2010 ; Vol. 21, No. 7. pp. 1321-1330.
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