Antibody organization on lipid films. Influence of pH and interchain disulphide reduction

Uzgiris, E. E.

doi:10.1042/bj2720045

Cited by 10 publications

(1 citation statement)

References 21 publications

(24 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“… 6–8 While the IgM molecules exist in a pentameric form, the IgG molecules exist in a monomeric form, but it was recently shown that the IgG molecules form a hexamer and the IgG hexamer binds to the complement complex by transmission electron microscopy. 9–11 Recently, we also reported the self-assembly of monoclonal IgG antibodies from mouse 12 using high-resolution atomic force microscopy (AFM) 13–20 ( Fig. 1b ).…”

Section: Introductionmentioning

confidence: 96%

Immunoactivity of self-assembled antibodies investigated by atomic force microscopy

et al. 2018

View full text Add to dashboard Cite

Immunoglobulin G (IgG), an antibody, plays a significant role in the immune system, and the functions of IgG molecules have been studied in many research fields such as medicine and engineering. Recently, we found the self-assembly of monoclonal mouse IgG molecules on a mica substrate using atomic force microscopy (AFM); the IgG molecules self-assemble into hexamers and the hexamers form a two-dimensional (2D) crystal. The self-assembly of the IgG molecules is of great interest in terms of the enhancement of the immunoactivity of the antibodies. In this study, we investigated the self-assembly of various IgG molecules on a mica substrate to discuss if the hexamerization of the IgG molecules is a general phenomenon. We also investigated the antigen binding site in the IgG antibody hexamers, and estimated the association rate constant of the self-assembled IgG molecules based on the AFM measurements.The estimated value was lower than that reported in a previous study probably because of the limited mobility of the antigen-binding fragments on the substrate.

show abstract

Section: Introductionmentioning

confidence: 96%

Immunoactivity of self-assembled antibodies investigated by atomic force microscopy

et al. 2018

View full text Add to dashboard Cite

show abstract

Inhibiting Nonspecific Protein Adsorption: Mechanisms, Methods, and Materials

Binazadeh

Zeng

Unsworth

2013

Biomaterials Surface Science

View full text Add to dashboard Cite

Synthesis of New Phospholipids Linked to Steroid‐Hormone Derivatives Designed for Two‐Dimensional Crystallization of Proteins

Lebeau

Oudet

Mioskowski

1991

Helvetica Chimica Acta

View full text Add to dashboard Cite

The synthesis of phospholipids 1n-3n, rationally designed for two-dimensional crystallization of progesterone and estrddiol receptors, is reported. The structure of these lipids provides them with essential properties such as fluidity and stability when spread into monolayers at the air/H20 interface, affinity for the protein to be crystallized, and accessibility of the ligand under the lipid monolayer.Introduction. -For a better understanding of enzymatic mechanisms and biological activities, knowledge of the three-dimensional (3D) structures of the macromolecules involved in these processes could be of value. Structural approaches using X-ray crystallography or nuclear magnetic resonance (NMR) need several milligrams (10-100 mg) of the highly purified protein (up to 95-98% pure). In the last ten years, a combination of two-dimensional (2D) crystallization and electron-microscopy techniques led to structure elucidation of proteins with resolution comparable to those reached by X-ray crystallography [l-141. This new approach requires only a few micrograms of partially purified biological material and allows the study of very high molecular weight proteins, whether they are soluble or not.2D Crystallization of proteins can be induced using lipid layers. The lipid, whose polar head is attached to a specific hgdnd of the protein, is allowed to spread at the air/H,O interface of a microvolume (typically 15 pl) of a buffered solution of the protein (Figs. l a and l b ) . Molecular recognition of the ligand by the protein results in placing the macromolecules in a plane with an orientation identical to that of its neighbours, except by rotation (axis perpendicular to the plane; F i g . 1~) . Due to the properties of the phospholipid (monolayer fluidity and stability) and those of the aqueous phase (ionic strength, pH, nature of ions in solution), protein-lipid complexes interacting one on each other (electrostatic interactions, H-bonds, hydrophobic insertions) move in rotation and translation to form 2D arrays of the protein (Fig. Id). These protein arrays are studied by electron microscopy, and after image processing, the 3D structure of the macromolecule can be obtained.In order to study the structure of steroid-hormone binding receptors, we synthesized molecules 1n-3fl whose structure confer to them specific physical and biological characteristics (Fig. 2). One of the most important points in the approach is the fluidity of the lipid film. Till now, all successfully performed experiments show that 2D crystals can be

show abstract

Antibody organization on lipid films. Influence of pH and interchain disulphide reduction

Cited by 10 publications

References 21 publications

Immunoactivity of self-assembled antibodies investigated by atomic force microscopy

Immunoactivity of self-assembled antibodies investigated by atomic force microscopy

Inhibiting Nonspecific Protein Adsorption: Mechanisms, Methods, and Materials

Synthesis of New Phospholipids Linked to Steroid‐Hormone Derivatives Designed for Two‐Dimensional Crystallization of Proteins

Contact Info

Product

Resources

About