Corrigendum: Advances in Plant-Derived Scaffold Proteins

AbstractProtein-based targeting reagents, such as antibodies and non-antibody scaffold proteins, are rapidly inactivated in the upper gastrointestinal (GI) tract. Hydrochloric acid in gastric juice denatures proteins and activates pepsin, concentrations of which reach 1 mg/mL in the mammalian stomach. Two stable scaffold proteins (nanobody and nanofitin), previously developed to be protease-resistant, were completely digested in less than 10 min at 100-fold lower concentration of pepsin than found in the stomach. Here we present gastrobodies, a protein scaffold derived from Kunitz soybean trypsin inhibitor (SBTI). SBTI is highly resistant to the challenges of the upper GI tract, including digestive proteases, pH 2 and bile acids. Computational prediction of SBTI’s evolvability identified two nearby loops for randomization, to create a potential recognition surface which was experimentally validated by alanine scanning. We established display of SBTI on full-length pIII of M13 phage. Phage selection of gastrobody libraries against the glucosyltransferase domain of Clostridium difficile toxin B (GTD) identified hits with nanomolar affinity and enzyme inhibitory activity. Anti-GTD binders retained high stability to acid, digestive proteases and heat. Gastrobodies show resilience to exceptionally harsh conditions, which should provide a foundation for targeting and modulating function within the GI tract.

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Variations in ADP-ribosylation of nuclear scaffold proteins during the Hela cell cycle

Biochemical and Biophysical Research Communications ◽

10.1016/0006-291x(85)90261-x ◽

1985 ◽

Vol 126 (2) ◽

pp. 840-847 ◽

Cited By ~ 14

Author(s):

Kenneth W. Adolph ◽

Min-Kyung H. Song

Keyword(s):

Cell Cycle ◽

Hela Cell ◽

Scaffold Proteins ◽

Nuclear Scaffold ◽

Adp Ribosylation

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Combinatorial Inhibition in PDZ Containing Scaffold Proteins

Biophysical Journal ◽

10.1016/j.bpj.2013.11.2692 ◽

2014 ◽

Vol 106 (2) ◽

pp. 476a

Author(s):

James McCann ◽

Ucheor B. Choi ◽

Mark E. Bowen

Keyword(s):

Scaffold Proteins

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p55 protein is a member of PSD scaffold proteins in the rat brain and interacts with various PSD proteins

Molecular Brain Research ◽

10.1016/j.molbrainres.2004.12.023 ◽

2005 ◽

Vol 135 (1-2) ◽

pp. 204-216 ◽

Cited By ~ 12

Author(s):

Zhang Jing-Ping ◽

Qing-Bao Tian ◽

Hiroyuki Sakagami ◽

Hisatake Kondo ◽

Shogo Endo ◽

...

Keyword(s):

Rat Brain ◽

Scaffold Proteins

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Differential Roles of JIP Scaffold Proteins in the Modulation of Amyloid Precursor Protein Metabolism

Journal of Biological Chemistry ◽

10.1074/jbc.m203713200 ◽

2002 ◽

Vol 277 (30) ◽

pp. 27567-27574 ◽

Cited By ~ 56

Author(s):

Hidenori Taru ◽

Yutaka Kirino ◽

Toshiharu Suzuki

Keyword(s):

Amyloid Precursor Protein ◽

Protein Metabolism ◽

Scaffold Proteins ◽

Precursor Protein ◽

Amyloid Precursor Protein Metabolism

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New interactions of invadopodia scaffold protein TKS5 with proteins that take part in actin cytoskeleton reorganization, endo-/exocytosis and membrane remodeling

Visnik ukrains'kogo tovaristva genetikiv i selekcioneriv ◽

10.7124/visnyk.utgis.16.2.1056 ◽

2019 ◽

Vol 16 (2) ◽

pp. 183-189

Author(s):

Y. M. Nemesh ◽

S. V. Kropyvko

Keyword(s):

Plasma Membrane ◽

Actin Cytoskeleton ◽

Scaffold Protein ◽

Scaffold Proteins ◽

Membrane Remodeling ◽

Sh3 Domains ◽

Protein Protein Interaction ◽

Cytoskeleton Reorganization ◽

New Interactions ◽

Cytoskeleton Rearrangement

Aim. TKS5 is a key scaffold protein of invadopodia. In its absence, the cells completely lose the ability to form invadopodia. This fact makes TKS5 a potential target for cancer cure and one of the central proteins in the investigation of cancer cell invasion. Additionally, the question remains about the function of TKS5 in normal cells. Therefore, in order to extend knowledge about TKS5 role in healthy and invasive cells, we tested the TKS5 interaction with the proteins involved in signal transduction: PLCγ1, SRC, CRK, CSK; the proteins involved in plasma membrane remodeling: AMPH1, BIN1, CIN85, ITSN1, ITSN2; the protein involved in the actin cytoskeleton rearrangement: CTTN. Methods. We used the GST Pull-down assay to identify the protein-protein interaction. Results. We revealed that TKS5 SH3 domains interact with CIN85. There were identified TKS5 interactions with SH3 domains of CTTN, ITSN1, ITSN2, AMPH1 and BIN1. Conclusions. TKS5 interacts with CIN85, CTTN, ITSN1, ITSN2, AMPH1 and BIN1, which take part in membrane remodeling, endo-/exocytosis and actin cytoskeleton rearrangement. Keywords: TKS5, scaffold proteins, actin cytoskeleton, plasma membrane.

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Role of Ezrin/Radixin/Moesin in the Surface Localization of Programmed Cell Death Ligand-1 in Human Colon Adenocarcinoma LS180 Cells

Pharmaceuticals ◽

10.3390/ph14090864 ◽

2021 ◽

Vol 14 (9) ◽

pp. 864

Author(s):

Takuro Kobori ◽

Chihiro Tanaka ◽

Mayuka Tameishi ◽

Yoko Urashima ◽

Takuya Ito ◽

...

Keyword(s):

Cell Death ◽

Plasma Membrane ◽

Programmed Cell Death ◽

Cell Surface ◽

Actin Cytoskeleton ◽

Mrna Level ◽

Scaffold Proteins ◽

Membrane Localization ◽

Erm Proteins ◽

Plasma Membrane Localization

Programmed cell death ligand-1 (PD-L1), an immune checkpoint protein highly expressed on the cell surface in various cancer cell types, binds to programmed cell death-1 (PD-1), leading to T-cell dysfunction and tumor survival. Despite clinical successes of PD-1/PD-L1 blockade therapies, patients with colorectal cancer (CRC) receive little benefit because most cases respond poorly. Because high PD-L1 expression is associated with immune evasion and poor prognosis in CRC patients, identifying potential modulators for the plasma membrane localization of PD-L1 may represent a novel therapeutic strategy for enhancing the efficacy of PD-1/PD-L1 blockade therapies. Here, we investigated whether PD-L1 expression in human colorectal adenocarcinoma cells (LS180) is affected by ezrin/radixin/moesin (ERM), functioning as scaffold proteins that crosslink plasma membrane proteins with the actin cytoskeleton. We observed colocalization of PD-L1 with all three ERM proteins in the plasma membrane and detected interactions involving PD-L1, the three ERM proteins, and the actin cytoskeleton. Furthermore, gene silencing of ezrin and radixin, but not of moesin, substantially decreased the expression of PD-L1 on the cell surface without affecting its mRNA level. Thus, in LS180 cells, ezrin and radixin may function as scaffold proteins mediating the plasma membrane localization of PD-L1, possibly by post-translational modification.

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