Английская Википедия:In situ cyclization of proteins

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Шаблон:Short description Шаблон:Cs1 config The in situ cyclization of proteins (INCYPRO) is a protein engineering technology that increases the durability of proteins and enzymes for biotechnological and biomedical applications.[1][2] For such applications, it is essential that the used proteins maintain their structural integrity.[3] This is, however, often challenged due to the conditions required for these applications which necessitates protein engineering to stabilize the protein structure.[4] The INCYPRO technology involves the attachment of molecular claps (crosslinks) to a protein, thereby reducing the tendency of the protein to unfold. The resulting INCYPRO-crosslinked proteins are more stable at elevated temperature and in presence of chemical denaturants.[5]

Technology

The INCYPRO technology utilizes tris-reactive molecules to crosslink three defined positions within a protein[1] or protein complex.[6] For example, INCYPRO can involve the introduction of three spatially aligned and solvent-accessible cysteines into the protein that are then reacted with a tris-electrophilic agent. The resulting crosslinked proteins or protein complexes have been shown to exhibit increased stability towards thermal and chemical stress and a lower tendency towards aggregation.[1][6] So far, the melting temperature of proteins was increased by up to 39°C in a single design step.[6]

Examples

An early example, involved the stabilization of the transpeptidase Sortase A which resulted in INCYPRO-stabilized variants with activity under elevated temperature and in the presence of guanidinium chloride.[1][5] INCYPRO has also been applied to stabilize the human adaptor KIX domain utilizing different crosslinker molecules. Here, a dependency of protein stability on the hydrophilicity of the crosslink was observed.[2] In addition, a number of homo-trimeric protein complexes was stabilized including the Pseudomonas fluorescens esterase (PFE) and an Enoyl-CoA hydratase.[6] In these cases, enzyme conjugates with overall bicyclic topology were generated.

See also

References

Шаблон:Reflist

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  1. ↑ 1,0 1,1 1,2 1,3 Шаблон:Cite journal
  2. ↑ 2,0 2,1 Шаблон:Cite journal
  3. ↑ Шаблон:Cite journal
  4. ↑ Шаблон:Cite journal
  5. ↑ 5,0 5,1 Шаблон:Cite journal
  6. ↑ 6,0 6,1 6,2 6,3 Шаблон:Cite journal
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