title: Kinetic analysis of his-tagged protein binding to nickel-chelating nanolipoprotein particles.
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.
authors.affiliation: Physical and Life Sciences Directorate, and Center for Accelerator Mass Spectrometry, Lawrence Livermore National Laboratory, Livermore, California 94551, USA.
authors.forename: Craig D
authors.initials: CD
authors.lastname: Blanchette
authors.forename: Nicholas O
authors.initials: NO
authors.lastname: Fischer
authors.forename: Michele
authors.initials: M
authors.lastname: Corzett
authors.forename: Graham
authors.initials: G
authors.lastname: Bench
authors.forename: Paul D
authors.initials: PD
authors.lastname: Hoeprich
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D002614:Chelating Agents / Q000737:chemistry* / Q000378:metabolism; D006639:Histidine / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008055:Lipids / Q000737:chemistry; D008074:Lipoproteins / Q000737:chemistry* / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry*; D009532:Nickel / Q000737:chemistry* / Q000378:metabolism; D010316:Particle Size; D011485:Protein Binding; D011994:Recombinant Proteins / Q000737:chemistry; D020349:Surface Plasmon Resonance; D015010:Yersinia pestis / Q000737:chemistry
chemical_list: D001426:Bacterial Proteins; D002614:Chelating Agents; D008055:Lipids; D008074:Lipoproteins; D011994:Recombinant Proteins; D006639:Histidine; D009532:Nickel