Immobilization of his-tagged proteins on nickel-chelating nanolipoprotein particles

Nicholas O Fischer, Craig D. Blanchette, Brett A. Chromy, Edward A. Kuhn, Brent W. Segelke, Michele Corzett, Graham Bench, Peter W. Mason, Paul D. Hoeprich

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Nanolipoprotein particles (NLPs) are nanometer-sized, discoidal particles that self-assemble from purified apolipoprotein and phospholipid. Their size and facile functionalization suggest potential application of NLPs as platforms for the presentation and delivery of recombinant proteins. To this end, we investigated incorporation of nickel-chelating lipids into NLPs (NiNLPs) and subsequent sequestration of polyhistidine (His)-tagged proteins. From initial lipid screens for NLP formation, the two phospholipids DMPC and DOPC were identified as suitable bulk lipids for incorporation of the nickel-chelating lipid DOGS-NTA-Ni into NLPs, and NiNLPs were successfully formed with varying amounts of DOGS-NTA-Ni. NiNLPs consisting of 10% DOGS-NTA-Ni with 90% bulk lipid (either DMPC or DOPC) were thoroughly characterized by size exclusion chromatography (SEC), non- denaturing gradient gel electrophoresis (NDGGE), and atomic force microscopy (AFM). Three different His- tagged proteins were sequestered on NiNLPs in a nickel-dependent manner, and the amount of immobilized protein was contingent on the size and composition of the NiNLP.

Original languageEnglish (US)
Pages (from-to)460-465
Number of pages6
JournalBioconjugate Chemistry
Volume20
Issue number3
DOIs
StatePublished - Mar 2009
Externally publishedYes

Fingerprint

Chelation
Nickel
Immobilization
Lipids
Proteins
Dimyristoylphosphatidylcholine
Phospholipids
Immobilized Proteins
Recombinant proteins
Denaturing Gradient Gel Electrophoresis
Size exclusion chromatography
Apolipoproteins
Atomic Force Microscopy
Electrophoresis
Recombinant Proteins
Gel Chromatography
Atomic force microscopy
Gels

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Organic Chemistry
  • Pharmaceutical Science
  • Biomedical Engineering
  • Pharmacology
  • Medicine(all)

Cite this

Fischer, N. O., Blanchette, C. D., Chromy, B. A., Kuhn, E. A., Segelke, B. W., Corzett, M., ... Hoeprich, P. D. (2009). Immobilization of his-tagged proteins on nickel-chelating nanolipoprotein particles. Bioconjugate Chemistry, 20(3), 460-465. https://doi.org/10.1021/bc8003155

Immobilization of his-tagged proteins on nickel-chelating nanolipoprotein particles. / Fischer, Nicholas O; Blanchette, Craig D.; Chromy, Brett A.; Kuhn, Edward A.; Segelke, Brent W.; Corzett, Michele; Bench, Graham; Mason, Peter W.; Hoeprich, Paul D.

In: Bioconjugate Chemistry, Vol. 20, No. 3, 03.2009, p. 460-465.

Research output: Contribution to journalArticle

Fischer, NO, Blanchette, CD, Chromy, BA, Kuhn, EA, Segelke, BW, Corzett, M, Bench, G, Mason, PW & Hoeprich, PD 2009, 'Immobilization of his-tagged proteins on nickel-chelating nanolipoprotein particles', Bioconjugate Chemistry, vol. 20, no. 3, pp. 460-465. https://doi.org/10.1021/bc8003155
Fischer, Nicholas O ; Blanchette, Craig D. ; Chromy, Brett A. ; Kuhn, Edward A. ; Segelke, Brent W. ; Corzett, Michele ; Bench, Graham ; Mason, Peter W. ; Hoeprich, Paul D. / Immobilization of his-tagged proteins on nickel-chelating nanolipoprotein particles. In: Bioconjugate Chemistry. 2009 ; Vol. 20, No. 3. pp. 460-465.
@article{15fb00472b014aed8be7bec97cbb0028,
title = "Immobilization of his-tagged proteins on nickel-chelating nanolipoprotein particles",
abstract = "Nanolipoprotein particles (NLPs) are nanometer-sized, discoidal particles that self-assemble from purified apolipoprotein and phospholipid. Their size and facile functionalization suggest potential application of NLPs as platforms for the presentation and delivery of recombinant proteins. To this end, we investigated incorporation of nickel-chelating lipids into NLPs (NiNLPs) and subsequent sequestration of polyhistidine (His)-tagged proteins. From initial lipid screens for NLP formation, the two phospholipids DMPC and DOPC were identified as suitable bulk lipids for incorporation of the nickel-chelating lipid DOGS-NTA-Ni into NLPs, and NiNLPs were successfully formed with varying amounts of DOGS-NTA-Ni. NiNLPs consisting of 10{\%} DOGS-NTA-Ni with 90{\%} bulk lipid (either DMPC or DOPC) were thoroughly characterized by size exclusion chromatography (SEC), non- denaturing gradient gel electrophoresis (NDGGE), and atomic force microscopy (AFM). Three different His- tagged proteins were sequestered on NiNLPs in a nickel-dependent manner, and the amount of immobilized protein was contingent on the size and composition of the NiNLP.",
author = "Fischer, {Nicholas O} and Blanchette, {Craig D.} and Chromy, {Brett A.} and Kuhn, {Edward A.} and Segelke, {Brent W.} and Michele Corzett and Graham Bench and Mason, {Peter W.} and Hoeprich, {Paul D.}",
year = "2009",
month = "3",
doi = "10.1021/bc8003155",
language = "English (US)",
volume = "20",
pages = "460--465",
journal = "Bioconjugate Chemistry",
issn = "1043-1802",
publisher = "American Chemical Society",
number = "3",

}

TY - JOUR

T1 - Immobilization of his-tagged proteins on nickel-chelating nanolipoprotein particles

AU - Fischer, Nicholas O

AU - Blanchette, Craig D.

AU - Chromy, Brett A.

AU - Kuhn, Edward A.

AU - Segelke, Brent W.

AU - Corzett, Michele

AU - Bench, Graham

AU - Mason, Peter W.

AU - Hoeprich, Paul D.

PY - 2009/3

Y1 - 2009/3

N2 - Nanolipoprotein particles (NLPs) are nanometer-sized, discoidal particles that self-assemble from purified apolipoprotein and phospholipid. Their size and facile functionalization suggest potential application of NLPs as platforms for the presentation and delivery of recombinant proteins. To this end, we investigated incorporation of nickel-chelating lipids into NLPs (NiNLPs) and subsequent sequestration of polyhistidine (His)-tagged proteins. From initial lipid screens for NLP formation, the two phospholipids DMPC and DOPC were identified as suitable bulk lipids for incorporation of the nickel-chelating lipid DOGS-NTA-Ni into NLPs, and NiNLPs were successfully formed with varying amounts of DOGS-NTA-Ni. NiNLPs consisting of 10% DOGS-NTA-Ni with 90% bulk lipid (either DMPC or DOPC) were thoroughly characterized by size exclusion chromatography (SEC), non- denaturing gradient gel electrophoresis (NDGGE), and atomic force microscopy (AFM). Three different His- tagged proteins were sequestered on NiNLPs in a nickel-dependent manner, and the amount of immobilized protein was contingent on the size and composition of the NiNLP.

AB - Nanolipoprotein particles (NLPs) are nanometer-sized, discoidal particles that self-assemble from purified apolipoprotein and phospholipid. Their size and facile functionalization suggest potential application of NLPs as platforms for the presentation and delivery of recombinant proteins. To this end, we investigated incorporation of nickel-chelating lipids into NLPs (NiNLPs) and subsequent sequestration of polyhistidine (His)-tagged proteins. From initial lipid screens for NLP formation, the two phospholipids DMPC and DOPC were identified as suitable bulk lipids for incorporation of the nickel-chelating lipid DOGS-NTA-Ni into NLPs, and NiNLPs were successfully formed with varying amounts of DOGS-NTA-Ni. NiNLPs consisting of 10% DOGS-NTA-Ni with 90% bulk lipid (either DMPC or DOPC) were thoroughly characterized by size exclusion chromatography (SEC), non- denaturing gradient gel electrophoresis (NDGGE), and atomic force microscopy (AFM). Three different His- tagged proteins were sequestered on NiNLPs in a nickel-dependent manner, and the amount of immobilized protein was contingent on the size and composition of the NiNLP.

UR - http://www.scopus.com/inward/record.url?scp=63749126132&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=63749126132&partnerID=8YFLogxK

U2 - 10.1021/bc8003155

DO - 10.1021/bc8003155

M3 - Article

C2 - 19239247

AN - SCOPUS:63749126132

VL - 20

SP - 460

EP - 465

JO - Bioconjugate Chemistry

JF - Bioconjugate Chemistry

SN - 1043-1802

IS - 3

ER -