Creation of artificial protein–protein interactions using α-helices as interfaces

Yagi, Sota; Akanuma, Satoshi; Yamagishi, Akihiko

doi:10.1007/s12551-017-0352-9

Cited by 9 publications

(8 citation statements)

References 70 publications

(100 reference statements)

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“…A second strategy fused the FLD of Angiopoietin 1 to the short CCOD of cartilage oligomeric matrix protein (COMP), which generated a pentameric complex with “super ligand” activity. However, while COMP‐ANG1 has improved solubility over native Angiopoietin 1, the coiled‐coil‐to‐coiled‐coil interactions, which tend to have broad interaction preference among different CCODs (Grigoryan & Keating, 2008; Yagi et al, 2018), have the propensity to trigger protein aggregation during production (Koh, 2013; Oh et al, 2015). It may also retain at the ECM after injection due to a general affinity of the CCOD towards tissue coiled‐coil proteins that are abundantly found in the ECM (Xu & Yu, 2001; Yamada & Sekiguchi, 2015).…”

Section: Discussionmentioning

confidence: 99%

New soluble angiopoietin analog of Hepta‐ANG1 prevents pathological vascular leakage

Liu

Ryczko²,

Xie

et al. 2020

Biotech & Bioengineering

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Vascular leak is a key driver of organ injury in diseases, and strategies that reduce enhanced permeability and vascular inflammation are promising therapeutic targets. Activation of the angiopoietin-1 (ANG1)-Tie2 tyrosine kinase signaling pathway is an important regulator of vascular quiescence. Here we describe the design and construction of a new soluble ANG1 mimetic that is a potent activator of endothelial Tie2 in vitro and in vivo. Using a chimeric fusion strategy, we replaced the extracellular matrix (ECM) binding and oligomerization domain of ANG1 with a heptameric scaffold derived from the C-terminus of serum complement protein C4binding protein α. We refer to this new fusion protein biologic as Hepta-ANG1, which forms a stable heptamer and induces Tie2 phosphorylation in cultured cells, and in the lung following intravenous injection of mice. Injection of Hepta-ANG1 ameliorates vascular endothelial growth factor-and lipopolysaccharide-induced vascular leakage, in keeping with the known functions of Angpt1-Tie2 in maintaining quiescent vascular stability. The new Hepta-ANG1 fusion is easy to produce and displays remarkable stability with high multimericity that can potently activate Tie2. It could be a new candidate ANG1 mimetic therapy for treatments of inflammatory vascular leak, such as acute respiratory distress syndrome and sepsis.

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Section: Discussionmentioning

confidence: 99%

New soluble angiopoietin analog of Hepta‐ANG1 prevents pathological vascular leakage

Liu

Ryczko²,

Xie

et al. 2020

Biotech & Bioengineering

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show abstract

“…A second strategy fused the FLD of ANGPT1 to the short coiled-coil domain of cartilage oligomeric matrix protein (COMP), which generated a pentameric complex with "super ligand" activity. However, while COMP-ANG1 has improved solubility over native ANG1, the coiledcoil-to-coiled-coil interactions, which tend to have broad interaction preference among different coiled-coil domains (32,33), have the propensity to trigger protein aggregation during production (15,25). It may also retain at the ECM after injection due to a general affinity of the coiled-coil domain towards tissue coiled-coil proteins that are abundantly found in the ECM (24,34).…”

Section: Discussionmentioning

confidence: 99%

New Soluble Angiopoietin Analog of C4BP-ANG1 Prevents Pathological Vascular Leakage

Liu

Ryczko²,

Xie

et al. 2020

Preprint

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Vascular leak is a key driver of organ injury in diseases such as Acute Respiratory Distress Syndrome caused by viruses, including COVID-19. Strategies that reduce enhanced permeability and vascular inflammation are promising therapeutic targets. Activation of the Angiopoietin-1 (Angpt1)-Tie2 tyrosine kinase signaling pathway is an important regulator of vascular quiescence. Here we describe the design and construction of a new soluble ANGPT1 mimetic that is a potent activator of endothelial Tie2 in vitro and in vivo. Using a chimeric fusion strategy, we replaced the extracellular matrix (ECM) binding and oligomerization domain of ANGPT1 with a heptameric scaffold derived from the C-terminus of serum complement protein C4-binding protein α (C4BP). We refer to this new fusion protein biologic as C4BP-ANG1, which forms a stable heptamer and induces TIE2 phosphorylation in cultured cells, and in the lung following i.v. injection of mice. Injection of C4BP-ANG1 ameliorates VEGF-and lipopolysaccharideinduced vascular leakage, in keeping with the known functions of Angpt1-Tie2 in maintaining quiescent vascular stability, and therefore is a promising candidate treatment for inflammatory endothelial dysfunction.

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“…An alternative approach for the rational design of de novo PPIs is the use of α-helices as the interfaces in de novo interactions. Here, one takes advantage of the well-known sequence–structure relationship of coiled coils, and indeed, this method has shown great promise (Figure B).…”

Section: Computational Tools For Engineering Protein–protein Interact...mentioning

confidence: 99%

Protein Engineering in the Design of Protein–Protein Interactions: SARS-CoV-2 Inhibitors as a Test Case

Zahradník

Schreiber

2021

Biochemistry

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The formation of specific protein−protein interactions (PPIs) drive most biological processes. Malfunction of such interactions is the molecular driver of many diseases. Our ability to engineer existing PPIs or create new ones has become a vital research tool. In addition, engineered proteins with new or altered interactions are among the most critical drugs that have been developed in recent years. These include antibodies, cytokines, inhibitors, and others. Here, we provide a perspective on the current status of the methods used to engineer new or altered PPIs. The emergence of the COVID-19 pandemic, which resulted in a worldwide quest to develop specific PPI inhibitors as drugs, provided an up-to-date and state-of-the-art status report on the methodologies for engineering PPIs targeting the interaction of the viral spike protein with its cellular target, ACE2. Multiple, very high affinity binders were generated within a few months using in vitro evolution by itself, or in combination with computational design. The different experimental and computational methods used to block this interaction provide a road map for the future of PPI engineering.

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Creation of artificial protein–protein interactions using α-helices as interfaces

Cited by 9 publications

References 70 publications

New soluble angiopoietin analog of Hepta‐ANG1 prevents pathological vascular leakage

New soluble angiopoietin analog of Hepta‐ANG1 prevents pathological vascular leakage

New Soluble Angiopoietin Analog of C4BP-ANG1 Prevents Pathological Vascular Leakage

Protein Engineering in the Design of Protein–Protein Interactions: SARS-CoV-2 Inhibitors as a Test Case

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