Non-specific clustering of histidine tagged green fluorescent protein mediated by surface interactions: the collective effect in the protein-adsorption behaviour

Abstract

The chemically programmed non-fouling surfaces were used to observe the surface assembly behaviour of hexahistidine tagged Green Fluorescent Protein (His-GFP) as a model protein. In this particular case, siteselective physisorption of His-GFP was achieved in the absence of metal ions. This preference does not arise from surface charge, wettability or topographic differences between regions. We found that His-GFP has a tendency to centre into an array of marked squares and acquires a template shape on the entire nonfouling surface when both the internal and surrounding areas present carboxylate groups. This surfacedirected organization of protein in assemblies is an unusual example of non-specific molecular interactions transfer to a higher scale objects organization. Furthermore, we performed a proof-of-concept study for the autonomous formation of protein microarrays with uniform orientation of the tagged protein molecules on the surface. Periodic protein microarrays were formed spontaneously within about one minute after the deposition of a few drops of protein solutions on the substrate. We propose a simple, gentle and cost-effective approach to fabrication of protein microarrays, which can be done by the enduser. This phenomenon of collective protein clustering into large scale patterns may help to assess experimentally how the peripheral proteins arrange into separate domains of the cell membrane.

Proteins Adsorbing onto Surface-Modified Nanoparticles: Effect of Surface Curvature, pH, and the Interplay of Polymers and Proteins Acid–Base Equilibrium

“…Future lines of work derive from the assumptions of the model employed in this work. Expanding our calculations from 1D to 3D systems (that is, without considering lateral homogeneity as we have done in this work) would allow us to study clustering of proteins, the role of surface properties, and whether this process is surface-mediated [77]. We would like to explore how protein changes upon adsorption can alter the process.…”

Tuning Electrostatic Interactions To Control Orientation of GFP Protein Adsorption on Silica Surface

“…13−15 In 2013, Sobiesćiak et alinvestigated the protein assemblies of His-tagged GFP proteins and found that the clustering of the proteins is a surface-mediated process, and that self-assembled monolayers (SAMs) can guide the formation of various protein arrays. 16 In 2017, Wasserberg et al examined the interaction between red fluorescence protein (RFP) tagged with histidines and Ni 2+ :nitrilotriacetic acid (NTA)-terminated thiols placed on an Au(111) slab. They discovered that the binding affinity increased with the length of the His tag, and the orientation could be precisely controlled by the careful placement of His tags.…”