Immobilization of histidine-tagged proteins on monodisperse metallochelation liposomes: Preparation and study of their structure
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
20732292
DOI
10.1016/j.ab.2010.08.023
PII: S0003-2697(10)00537-3
Knihovny.cz E-resources
- MeSH
- Chelating Agents chemistry MeSH
- Histidine chemistry genetics metabolism MeSH
- HIV Envelope Protein gp120 chemistry genetics metabolism MeSH
- HIV-1 metabolism MeSH
- Immobilized Proteins chemistry genetics metabolism MeSH
- Humans MeSH
- Liposomes chemistry MeSH
- Micelles MeSH
- Microscopy, Atomic Force MeSH
- Nickel chemistry MeSH
- Oligopeptides chemistry genetics metabolism MeSH
- Proteolipids chemistry MeSH
- Microscopy, Electron, Transmission MeSH
- Ultrafiltration methods MeSH
- Green Fluorescent Proteins chemistry genetics metabolism MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Chelating Agents MeSH
- gp120 protein, Human immunodeficiency virus 1 MeSH Browser
- His-His-His-His-His-His MeSH Browser
- Histidine MeSH
- HIV Envelope Protein gp120 MeSH
- Immobilized Proteins MeSH
- Liposomes MeSH
- Micelles MeSH
- Nickel MeSH
- Oligopeptides MeSH
- Proteolipids MeSH
- proteoliposomes MeSH Browser
- Green Fluorescent Proteins MeSH
Liposomes represent a biocompatible platform for the construction of self-assembling proteoliposomes using nickel or zinc metallochelation. Potential applications of such structures consist in the development of new biocompatible vaccination nanoparticles and drug delivery nanoparticle systems. Here, we describe the design and construction of a flow-through ultrafiltration cell suitable for the preparation of monodisperse liposomes enabled for metallochelation and, hence, the formation of proteoliposomes. The linkage of the cell with a fast protein liquid chromatography system facilitates automation of the procedure, which fits the criteria for upscaling. Proof-of-concept experiments are performed using a mixture of egg phosphatidyl choline and nickel-chelating lipid DOGS-NTA-Ni (1,2-dioleoyl-sn-glycero-3-{[N(5-amino-1-carboxypentyl)iminodiacetic acid]succinyl}(nickel salt)) to formulate proteoliposomes with proteins attached by metallochelation, including histidine (His)-tagged recombinant green fluorescent protein and rgp120 (derived from HIV-1 Env). These model proteoliposomes are characterized by gel permeation chromatography and by dynamic light scattering. Transmission electron microscopy and immunogold staining are used to characterize surface-bound proteins, revealing the tendency of rgp120 to form microdomains on liposome surfaces. These microdomains possess a two-dimensional crystal-like structure that is seen more precisely by atomic force microscopy.
References provided by Crossref.org
Targeting Human Thrombus by Liposomes Modified with Anti-Fibrin Protein Binders
