Deciphering the Stepwise Binding Mode of HRG1β to HER3 by Surface Plasmon Resonance and Interaction Map

Peeß, Carmen; Proff, Leopold von; Goller, Sabine; Andersson, Karl; Gerg, Michael; Malmqvist, Magnus; Bossenmaier, Birgit; Schräml, Michael

doi:10.1371/journal.pone.0116870

Cited by 10 publications

(5 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2C). Binding of lumretuzumab to HER3 blocks HER2/HER3 dimer formation and prevents ligand-dependent phosphorylation of both receptors (5,26), as also observed in Caco2 cells exposed to lumretuzumab (Fig. 2C).…”

Section: Cellular Model For Mechanistic Investigations Of Gastrointessupporting

confidence: 69%

Mechanistic Investigations of Diarrhea Toxicity Induced by Anti-HER2/3 Combination Therapy

Moisan

Michielin

Jacob

et al. 2018

Molecular Cancer Therapeutics

View full text Add to dashboard Cite

Combination of targeted therapies is expected to provide superior efficacy in the treatment of cancer either by enhanced antitumor activity or by preventing or delaying the development of resistance. Common challenges in developing combination therapies include the potential of additive and aggravated toxicities associated with pharmacologically related adverse effects. We have recently reported that combination of anti-HER2 and anti-HER3 antibodies, pertuzumab and lumretuzumab, along with paclitaxel chemotherapy in metastatic breast cancer, resulted in a high incidence of diarrhea that ultimately limited further clinical development of this combination. Here, we further dissected the diarrhea profile of the various patient dose cohorts and carried out investigations in human colon cell lines and explants to decipher the contribution and the mechanism of anti-HER2/3 therapeutic antibodies to intestinal epithelium malfunction. Our clinical investigations in patients revealed that while dose reduction of lumretuzumab, omission of pertuzumab loading dose, and introduction of a prophylactic antidiarrheal treatment reduced most severe adverse events, patients still suffered from persistent diarrhea during the treatment. Our investigations showed that pertuzumab and lumretuzumab combination treatment resulted in upregulation of chloride channel activity without indication of intestinal barrier disruption. Overall, our findings provide a mechanistic rationale to explore alternative of conventional antigut motility using medication targeting chloride channel activity to mitigate diarrhea of HER combination therapies. .

show abstract

Section: Cellular Model For Mechanistic Investigations Of Gastrointessupporting

confidence: 69%

Mechanistic Investigations of Diarrhea Toxicity Induced by Anti-HER2/3 Combination Therapy

Moisan

Michielin

Jacob

et al. 2018

Molecular Cancer Therapeutics

View full text Add to dashboard Cite

show abstract

“…Purified RcFLS1-His was injected in a concentration series of 1475 nM, 737.5 nM, 368.8 nM, 184.4 nM, and 92.2 nM RcFLS2-His was injected in a similar concentration series in another assay. Parameter settings and specific measurement methods were described previously ( Peess et al., 2015 ). For the split-luciferase (LUC) complementation assays, pCAMBIA1300-nLUC and pCAMBIA1300-cLUC vectors were selected.…”

Section: Methodsmentioning

confidence: 99%

Competition between FLS and DFR regulates the distribution of flavonols and proanthocyanidins in Rubus chingii Hu

Lei

Huang²,

Ruan³

et al. 2023

Front. Plant Sci.

View full text Add to dashboard Cite

Rubus chingii Hu is a berry plant of the genus Rubus of the Rosaceae family, which has high nutritional and medicinal value and is rich in flavonoids. Flavonol synthase (FLS) and dihydroflavonol 4-reductase (DFR) compete for the common substrate dihydroflavonols to regulate the metabolic flux of flavonoids. However, the competition between FLS and DFR based on enzyme is rarely reported. Here, we isolated and identified two FLS genes (RcFLS1 and RcFLS2) and one DFR gene (RcDFR) from Rubus chingii Hu. RcFLSs and RcDFR were highly expressed in stems, leaves, and flowers, although the flavonol accumulation in these organs was significantly higher than that of proanthocyanidins (PAs). The recombinant RcFLSs demonstrated bifunctional activities via hydroxylation and desaturation at the C-3α position having a lower Michaelis constant (Km) for dihydroflavonols than RcDFR. We also found that a low concentration of flavonols could significantly inhibit RcDFR activity. To investigate the competitive relationship between RcFLSs and RcDFR, we used a prokaryotic expression system (E. coli) to co-express these proteins. The transgenic cells expressing recombinant proteins were incubated with substrates, and the reaction products were analyzed. Furthermore, two transient expression systems (tobacco leaves and strawberry fruits) and a stable genetic system (Arabidopsis thaliana) were used to co-express these proteins in vivo. The results showed that RcFLS1 was dominant in the competition with RcDFR. Our results demonstrated that the competition between FLS and DFR regulated the metabolic flux distribution of flavonols and PAs, which will be of great significance for the molecular breeding of Rubus plants.

show abstract

“…There have also been reports of affixing multiple ligands on a sensor surface in a solution-like context using DNA-directed immobilization using SPR, which would be useful for characterizing BsAbs. New biosensor technologies that allow for discrete detection of both binding events and precise control over surface density, as well as advances in SPR data analysis and experimental design, may provide avenues for more thoroughly investigating complex binding events under increasingly biologically relevant conditions in the future [105][106][107][108][109].…”

Section: Bsab Bioassay: Binding Assaysmentioning

confidence: 99%

Bioassay Development for Bispecific Antibodies—Challenges and Opportunities

Tarighat

Lee

2021

IJMS

View full text Add to dashboard Cite

Antibody therapeutics are expanding with promising clinical outcomes, and diverse formats of antibodies are further developed and available for patients of the most challenging disease areas. Bispecific antibodies (BsAbs) have several significant advantages over monospecific antibodies by engaging two antigen targets. Due to the complicated mechanism of action, diverse structural variations, and dual-target binding, developing bioassays and other types of assays to characterize BsAbs is challenging. Developing bioassays for BsAbs requires a good understanding of the mechanism of action of the molecule, principles and applications of different bioanalytical methods, and phase-appropriate considerations per regulatory guidelines. Here, we review recent advances and case studies to provide strategies and insights for bioassay development for different types of bispecific molecules.

show abstract

Deciphering the Stepwise Binding Mode of HRG1β to HER3 by Surface Plasmon Resonance and Interaction Map

Cited by 10 publications

References 28 publications

Mechanistic Investigations of Diarrhea Toxicity Induced by Anti-HER2/3 Combination Therapy

Mechanistic Investigations of Diarrhea Toxicity Induced by Anti-HER2/3 Combination Therapy

Competition between FLS and DFR regulates the distribution of flavonols and proanthocyanidins in Rubus chingii Hu

Bioassay Development for Bispecific Antibodies—Challenges and Opportunities

Contact Info

Product

Resources

About