Factors affecting the physical stability (aggregation) of peptide therapeutics

Zapadka, Karolina L.; Becher, Frederik J.; Santos, Ana Lúcia Gomes dos; Jackson, Sophie

doi:10.1098/rsfs.2017.0030

Cited by 252 publications

(233 citation statements)

References 242 publications

(309 reference statements)

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“…Hence, the decoding capacity of our heterologous expression system might cause the cells' inability to distinguish the source of these ligands, leading to so-called "system bias". Our data suggests that peptide-based FPR1 pharmacotherapy might be worthwhile exploring, however, peptide aggregation is a huge challenge [59].…”

Section: Discussionmentioning

confidence: 92%

Exploring Biased Agonism at FPR1 as a Means to Encode Danger Sensing

Gröper

König

Kostenis

et al. 2020

Cells

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Ligand-based selectivity in signal transduction (biased signaling) is an emerging field of G protein-coupled receptor (GPCR) research and might allow the development of drugs with targeted activation profiles. Human formyl peptide receptor 1 (FPR1) is a GPCR that detects potentially hazardous states characterized by the appearance of N-formylated peptides that originate from either bacteria or mitochondria during tissue destruction; however, the receptor also responds to several non-formylated agonists from various sources. We hypothesized that an additional layer of FPR signaling is encoded by biased agonism, thus allowing the discrimination of the source of threat. We resorted to the comparative analysis of FPR1 agonist-evoked responses across three prototypical GPCR signaling pathways, i.e., the inhibition of cAMP formation, receptor internalization, and ERK activation, and analyzed cellular responses elicited by several bacteria-and mitochondria-derived ligands. We also included the anti-inflammatory annexinA1 peptide Ac2-26 and two synthetic ligands, the W-peptide and the small molecule FPRA14. Compared to the endogenous agonists, the bacterial agonists displayed significantly higher potencies and efficacies. Selective pathway activation was not observed, as both groups were similarly biased towards the inhibition of cAMP formation. The general agonist bias in FPR1 signaling suggests a source-independent pathway selectivity for transmission of pro-inflammatory danger signaling.Cells 2020, 9, 1054 2 of 18 translation. However, mitochondria, which are considered to represent endosymbionts of bacterial origin [3], initiate protein biosynthesis with N-formylated methionine and might release detectable levels of formylated peptides in situations of enhanced cell death or trauma [4,5]. Therefore, the appearance of N-formylated peptides signals potentially hazardous states caused by either bacterial threats (PAMPs) or tissue destruction (DAMPs).In higher eukaryotes, this unique pattern is sensed by the family of formyl peptide receptors (FPRs), which belong to the superfamily of G protein-coupled receptors (GPCRs) [5,6]. Thus, a shared sensor system detects bacteria-derived ligands and those that are liberated by non-infectious tissue destruction from mitochondria. The corresponding formylated peptides elicit strong signals via the activation of formyl peptide receptor 1 (FPR1) [7,8]. Here, we addressed whether human FPR1 is capable to decode the actual source of threat via different, i.e., ligand-specific signaling, and consequently modify the elicited cellular responses. We hypothesized that these layers of signal information are encoded by distinguishing ligand perception and potentially are governed by biased agonism. This emerging concept in GPCR-mediated signal transduction [9] emphasizes that ligands selectively stabilize specific receptor conformations that ultimately favor one (or more) signaling pathways out of the many the receptor is linked to. The final cellular response elicited by the receptor/ligand...

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Section: Discussionmentioning

confidence: 92%

Exploring Biased Agonism at FPR1 as a Means to Encode Danger Sensing

Gröper

König

Kostenis

et al. 2020

Cells

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“…At the same time, as the biopharmaceutical market expands, the need of recombinant proteins increases. For that reason, most of the factors affecting physical stability (aggregation) of peptides have been analyzed [23]. In addition, several methods based on algorithms are used for the prediction of the aggregation tendency of peptides and proteins [8,24] in order to reduce or avoid formation of intermolecular interactions.…”

Section: Modulation Of Recombinant Protein Solubility In Clearcoli Cellsmentioning

confidence: 99%

Insoluble proteins catch heterologous soluble proteins into inclusion bodies by intermolecular interaction of aggregating peptides

Carratalá

Cisneros

Hellman

et al. 2020

Preprint

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Background Protein aggregation is a biological event observed in expression systems in which the recombinant protein is produced under stressful conditions surpassing the homeostasis of the protein quality control system. In addition, protein aggregation is also related to conformational diseases in animals as transmissible prion diseases or non-transmissible neurodegenerative diseases including Alzheimer, Parkinson's disease, amyloidosis and multiple system atrophy among others. At the molecular level, the presence of aggregating-prone domains in protein molecules act as seeding igniters to induce the accumulation of protein molecules in protease-resistant clusters by intermolecular interactions. Results In this work we have studied the aggregating-prone performance of a small peptide (L6K2) with additional antimicrobial activity and we have elucidated the relevance of the accompanying scaffold protein to enhance the aggregating profile of the fusion protein. Furthermore, we demonstrated that the fusion of L6K2 to highly soluble recombinant proteins directs the protein to inclusion bodies (IB) in E. coli through stereospecific interactions in the presence of an insoluble protein displaying the same aggregating-prone peptide (APP). Conclusions These data suggest that the molecular bases of protein aggregation are related to the net balance of protein aggregation potential and not only to the presence of aggregation-prone stretches. This is then presented as a generic platform to generate hybrid protein aggregates in microbial cell factories for biopharmaceutical and biotechnological applications.

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“…Aggregation represents a significant limitation to the overall promise of peptide therapeutics as the formation of aggregates can limit their potency and lead to immunogenicity . Aggregates can be disordered globules or they can be highly ordered structures . Although highly ordered aggregates may also be considered self‐assembled structures, we distinguish aggregation processes as those being undesired for the peptide of interest.…”

Section: Protein Engineering To Design Peptide Characteristicsmentioning

confidence: 99%

Beyond function: Engineering improved peptides for therapeutic applications

2019

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Peptides are a promising source of new therapeutics, but the biophysical characteristics of natural peptides, including their stability and propensity to aggregate, can limit their success. Protein engineering offers powerful tools to improve the properties of peptides for biological applications. In this review, we explain rational design, directed evolution, and computational methods and how these methods can be applied to improving the characteristics of peptides. We also provide a discussion of engineering the thermodynamic stability, self‐assembly, reduced aggregation, proteolytic stability, and binding affinity and specificity of peptides, along with a perspective on future directions in engineering therapeutic peptides.

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Factors affecting the physical stability (aggregation) of peptide therapeutics

Cited by 252 publications

References 242 publications

Exploring Biased Agonism at FPR1 as a Means to Encode Danger Sensing

Exploring Biased Agonism at FPR1 as a Means to Encode Danger Sensing

Insoluble proteins catch heterologous soluble proteins into inclusion bodies by intermolecular interaction of aggregating peptides

Beyond function: Engineering improved peptides for therapeutic applications

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