Abstract
Histidine-rich peptides are commonly used in recombinant protein production as purification tags, allowing the one-step affinity separation of the His-tagged proteins from the extracellular media or cell extracts. Genetic engineering makes feasible the post-purification His-tag removal by inserting, between the tag and the main protein body, a target site for trans-acting proteases or a self-proteolytic peptide with regulatable activities. However, for technical ease, His tags are often not removed and the fusion proteins eventually used in this form. In this commentary, we revise the powerful biological properties of histidine-rich peptides as endosomolytic agents and as architectonic tags in nanoparticle formation, for which they are exploited in drug delivery and other nanomedical applications. These activities, generally unknown to biotechnologists, can unwillingly modulate the functionality and biotechnological performance of recombinant proteins in which they remain trivially attached.
Highlights
Histidine-rich peptides are commonly used in recombinant protein production as purification tags, allowing the one-step affinity separation of the His-tagged proteins from the extracellular media or cell extracts
His-tags do not add significant metabolic load to the protein production process and they can be incorporated to the protein by simple genetic engineering at the upstream level
Protein production scientists are usually unaware of the potent biological properties of histidine-rich peptides for which they are used in nanomedicine, which are activated at acidic pH
Summary
Protein production scientists are usually unaware of the potent biological properties of histidine-rich peptides for which they are used in nanomedicine (essentially endosomal disruption and cross-molecular electrostatic interactions), which are activated at acidic pH. The same (or very similar) His-rich peptides used in affinity chromatography, incorporated into different type of nanoparticles (most ranging from H2 to H10, Table 2) to favour intracellular trafficking in drug delivery [25], are employed as affinity tags for separation and often not removed after protein purification (Table 1). These remaining agents might render unexpected protein behaviour, when exposed to cells or nucleic acids, and specific measures for control should be implemented in biological assays when technical obstacles make His-tag removal inconvenient upon protein purification
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