The autoinhibitory function of D1 domain of FGFR1 goes beyond the inhibition of ligand binding

Opaliński, Łukasz; Szczepara, Martyna; Sokołowska-Wędzina, Aleksandra; Zakrzewska, Małgorzata; Otlewski, Jacek

doi:10.1016/j.biocel.2017.06.015

Cited by 12 publications

(15 citation statements)

References 15 publications

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“…We employed a biochemical assay where we blocked the synthesis of new FGFR1 molecules with cycloheximide and analyzed with western blotting the levels of FGFR1 in time upon stimulation with B‐Fc and T‐Fc. The time‐dependent decrease in FGFR1 level is attributed to the lysosomal degradation of the internalized receptor [5,6]. In nontreated cells within the time of the experiment, we observed very slight reduction in FGFR1 level that likely corresponds to the constitutive, ligand‐independent receptor endocytosis (Fig.…”

Section: Resultsmentioning

confidence: 81%

“…We have recently reported a phage display‐based selection of high‐affinity antibody fragments that selectively recognize epitopes within the D1 domain of the FGFR1 [25]. Furthermore, we have demonstrated that the bivalency and the high affinity promote internalization of FGFR1/engineered antibody complexes [5,6,49]. To study the impact of FGFR1 clustering into larger oligomeric structures on the receptor activity and intracellular trafficking, we generated tetravalent engineered antibody, T‐Fc, composed of two anti‐FGFR1 scFv fragments recognizing D1 domain of the receptor fused to the Fc fragment of human IgG1.…”

Section: Resultsmentioning

confidence: 99%

“…Fully glycosylated extracellular domains of FGFRs fused to the Fc fragment of human IgG1: FGFR1 IIIc (FGFR1‐Fc), FGFR2 IIIc (FGFR2‐Fc), FGFR3 IIIc (FGFR3‐Fc), and FGFR4 (FGFR4‐Fc), and the extracellular part of FGFR1 lacking the N‐terminal D1 domain (FGFR1 D2‐D3) were produced as described previously by our group [48]. FGFR1 GST‐D1 was expressed in E. coli BL21 CodonPlus (DE3)‐RIL (Agilent Technologies, Santa Clara, CA, USA) and purified with use of Glutathione Sepharose column as described previously [5,6,49].…”

Section: Methodsmentioning

confidence: 99%

“…The extracellular domain of FGFR1 contains three immunoglobulin‐like domains D1, D2, and D3. The D1 domain fulfills a regulatory function preventing FGFR1 from autoactivation in the absence of FGFs [3–6]. The D2 and D3 domains form FGF binding sites [1].…”

Section: Introductionmentioning

confidence: 99%

“…The uptake of FGF/FGFR1 complexes mainly occurs via clathrin‐mediated endocytosis (CME) leading to lysosomal degradation of FGFR1 [29,30,36,37,39]. We have recently uncoupled FGFR1 dimerization from the receptor activation and have demonstrated that dimerization of FGFR1, but not receptor autophosphorylation, triggers CME of FGFR1 [5,6]. A number of proteins involved in CME of FGFR1 in response to FGF binding were identified,however, the exact role of a number of these factors in the receptor internalization is still largely mysterious [31,40–44].…”

Section: Introductionmentioning

confidence: 99%

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FGFR1 clustering with engineered tetravalent antibody improves the efficiency and modifies the mechanism of receptor internalization

et al. 2020

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Fibroblast growth factor receptor 1 (FGFR1) transmits signals through the plasma membrane regulating essential cellular processes like division, motility, metabolism, and death. Overexpression of FGFR1 is observed in numerous tumors and thus constitutes an attractive molecular target for selective cancer treatment. Targeted anti‐cancer therapies aim for the precise delivery of drugs into cancer cells, sparing the healthy ones and thus limiting unwanted side effects. One of the key steps in targeted drug delivery is receptor‐mediated endocytosis. Here, we show that the efficiency and the mechanism of FGFR1 internalization are governed by the spatial distribution of the receptor in the plasma membrane. Using engineered antibodies of different valency, we demonstrate that dimerization of FGFR1 with bivalent antibody triggers clathrin‐mediated endocytosis (CME) of the receptor. Clustering of FGFR1 into larger oligomers with tetravalent antibody stimulates fast and highly efficient uptake of the receptor that occurs via two distinct mechanisms: CME and dynamin‐dependent clathrin‐independent endocytic routes. Furthermore, we show that all endocytic pathways engaged in FGFR1 internalization do not require receptor activation. Our data provide novel insights into the mechanisms of intracellular trafficking of FGFR1 and constitute guidelines for development of highly internalizing antibody‐based drug carriers for targeted therapy of FGFR1‐overproducing cancers.

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Section: Resultsmentioning

confidence: 81%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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FGFR1 clustering with engineered tetravalent antibody improves the efficiency and modifies the mechanism of receptor internalization

et al. 2020

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New developments in the biology of fibroblast growth factors

Ornitz

Itoh

2022

WIREs Mechanisms of Disease

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The fibroblast growth factor (FGF) family is composed of 18 secreted signaling proteins consisting of canonical FGFs and endocrine FGFs that activate four receptor tyrosine kinases (FGFRs 1-4) and four intracellular proteins (intracellular FGFs or iFGFs) that primarily function to regulate the activity of voltagegated sodium channels and other molecules. The canonical FGFs, endocrine FGFs, and iFGFs have been reviewed extensively by us and others. In this review, we briefly summarize past reviews and then focus on new developments in the FGF field since our last review in 2015. Some of the highlights in the past 6 years include the use of optogenetic tools, viral vectors, and inducible transgenes to experimentally modulate FGF signaling, the clinical use of small molecule FGFR inhibitors, an expanded understanding of endocrine FGF signaling, functions for FGF signaling in stem cell pluripotency and differentiation, roles for FGF signaling in tissue homeostasis and regeneration, a continuing elaboration of mechanisms of FGF signaling in development, and an expanding appreciation of roles for FGF signaling in neuropsychiatric diseases.

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Glycosylation of FGF/FGFR: An underrated sweet code regulating cellular signaling programs

Gędaj,

Gregorczyk,

Żukowska

et al. 2024

Cytokine & Growth Factor Reviews

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The autoinhibitory function of D1 domain of FGFR1 goes beyond the inhibition of ligand binding

Cited by 12 publications

References 15 publications

FGFR1 clustering with engineered tetravalent antibody improves the efficiency and modifies the mechanism of receptor internalization

FGFR1 clustering with engineered tetravalent antibody improves the efficiency and modifies the mechanism of receptor internalization

New developments in the biology of fibroblast growth factors

Glycosylation of FGF/FGFR: An underrated sweet code regulating cellular signaling programs

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