Lecture

Circular Permutation in Protein Engineering

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In course

CH-319: Experimental biochemistry and biophysics

A 7-week long (4+8 h) experiment where you plan and construct a fluorescent sensor protein starting from DNA bricks. The protein will be expressed in and purified from E.coli, characterized by biochemical and spectroscopic methods. Your report will be in ...

Description

This lecture explores circular permutation in protein engineering, a technique used to optimize proteins by introducing new N- and C-termini without altering the protein structure. The instructor discusses the concept of circular permutation, the process of creating new termini, and the importance of linkers in maintaining protein functionality. Additionally, the lecture covers the review of FRET parameters and the application of circular permutation in FRET optimization. Examples of circular permutation in genetically encoded fluorescent indicators and voltage biosensors are also presented.

Instructor

Rudolf Hovius

Expertise Â•Protein structure, expression & purificationÂ•Molecular interaction & receptor activationÂ•Fluorescence imaging & spectroscopySignalling & sensing Mission => Teaching: Lecture courses, tutorials and practicals in biochemistry, biophysical chemistry of membranes and signalling processes. Participating to the Commission d'Enseignement and Commission de l'Ecole Doctorale of Chemistry and Chemical Engineering.=> Research interests: Protein function and structure, Membrane protein (especially recepto

Official source

Related concepts (16)

Protein engineering

Protein engineering is the process of developing useful or valuable proteins through the design and production of unnatural polypeptides, often by altering amino acid sequences found in nature. It is a young discipline, with much research taking place into the understanding of protein folding and recognition for protein design principles. It has been used to improve the function of many enzymes for industrial catalysis. It is also a product and services market, with an estimated value of $168 billion by 2017.

Green fluorescent protein

The green fluorescent protein (GFP) is a protein that exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range. The label GFP traditionally refers to the protein first isolated from the jellyfish Aequorea victoria and is sometimes called avGFP. However, GFPs have been found in other organisms including corals, sea anemones, zoanithids, copepods and lancelets. The GFP from A. victoria has a major excitation peak at a wavelength of 395 nm and a minor one at 475 nm.

Förster resonance energy transfer

Förster resonance energy transfer (FRET), fluorescence resonance energy transfer, resonance energy transfer (RET) or electronic energy transfer (EET) is a mechanism describing energy transfer between two light-sensitive molecules (chromophores). A donor chromophore, initially in its electronic excited state, may transfer energy to an acceptor chromophore through nonradiative dipole–dipole coupling. The efficiency of this energy transfer is inversely proportional to the sixth power of the distance between donor and acceptor, making FRET extremely sensitive to small changes in distance.

Protein design

Protein design is the rational design of new protein molecules to design novel activity, behavior, or purpose, and to advance basic understanding of protein function. Proteins can be designed from scratch (de novo design) or by making calculated variants of a known protein structure and its sequence (termed protein redesign). Rational protein design approaches make protein-sequence predictions that will fold to specific structures.

Genetic engineering

Genetic engineering, also called genetic modification or genetic manipulation, is the modification and manipulation of an organism's genes using technology. It is a set of technologies used to change the genetic makeup of cells, including the transfer of genes within and across species boundaries to produce improved or novel organisms. New DNA is obtained by either isolating and copying the genetic material of interest using recombinant DNA methods or by artificially synthesising the DNA.

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