Recombinant expression a novel fibronectin—collage fusion peptide modulating stem cell stemness via integrin β3

Luo, Xin; Geng, Dezhi; Zhang, Qirong; Ye, Tao; Zhang, Yifan; Li, Ziyi; Huang, Yadong; Xiang, Qi

doi:10.1007/s00253-022-11965-4

Cited by 7 publications

(3 citation statements)

References 40 publications

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“…After obtaining high-purity 0630 protein, the characterization of the secondary structure of 0630 protein in solution was carried out by CD spectroscopy, which evaluates the secondary structure by using the differential absorption of left- and right-handed circularly polarized light in asymmetric environments . The results were obtained with a very pronounced negative absorption peak at 198 nm, but a small and broad positive absorption peak at 250–300 nm This may be due to the presence of irregular coil structures in the protein secondary structure, suggesting that the spectral characteristics of the 0630 protein are similar to those of typical denatured collagen. , These results suggest that the 0630 structure is predominantly random coils.…”

Section: Discussionmentioning

confidence: 99%

Production and Functional Analysis of Collagen Hexapeptide Repeat Sequences in Pichia pastoris

Guo,

Wang,

Yuwen

et al. 2024

J. Agric. Food Chem.

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In the development of biomaterials with specific structural domains associated with various cellular activities, the limited integrin specificity of commonly used adhesion sequences, such as the RGD tripeptide, has resulted in an inability to precisely control cellular responses. To overcome this limitation, we conducted multiple replications of the integrin α 2 β 1 -specific ligand, the collagen hexapeptide Gly-Phe-Pro-Gly-Glu-Arg (GFPGER) in Pichia pastoris. This enabled the development of recombinant collagen with high biological activity, which was subsequently expressed, isolated, and purified for structural and functional analysis. The proteins carrying the multiple replications GFPGER sequence demonstrated significant bioactivity in cells, leading to enhanced cell adhesion, osteoblast differentiation, and mineralization when compared to control groups. Importantly, these effects were mediated by integrin α 2 β 1 . The new collagen constructed in this study is expected to be a biomaterial for regulating specific cell functions and fates.

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Section: Discussionmentioning

confidence: 99%

Production and Functional Analysis of Collagen Hexapeptide Repeat Sequences in Pichia pastoris

Guo,

Wang,

Yuwen

et al. 2024

J. Agric. Food Chem.

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“…Previous studies indicated that fibronectin can be subdivided into plasma fibronectin and cellular fibronectin, the former one is always synthesized by hepatocytes and the latter one is produced by a variety of cells, such as chondrocytes and so on [ 14 ]. Furthermore, fibronectin is well-documented for its function in several biological procedures containing cell adhesion, migration, stemness, differentiation and angiogenesis [ 15 – 18 ]. A lot of studies have indicated that cell adhesion to extracellular fibronectin played a key role in tumorigenesis[ 19 ], and malignant progression in multiple cancer types, such as the development of the resistance to anoikis and chemotherapy [ 20 , 21 ], cancer metastasis [ 22 ], and so on.…”

Section: Introductionmentioning

confidence: 99%

Fibronectin promotes tumor angiogenesis and progression of non-small-cell lung cancer by elevating WISP3 expression via FAK/MAPK/ HIF-1α axis and activating wnt signaling pathway

Zhou

Sun

et al. 2023

Exp Hematol Oncol

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Background Fibronectin, an extracellular matrix protein, has been reported to be associated with heterogeneous cancer stemness, angiogenesis and progression in multiple cancer types. However, the roles and the underlying mechanism of fibronectin on the progression NSCLC need to be further elucidated. Methods Public dataset such as Kaplan-Meier Plotter was used to determine the prognostic significance of genes. The correlation of different protein expression in clinical and xenograft tissues was tested by immunohistochemistry experiment. Both in vitro and in vivo experiments were performed to determine the role of fibronectin on the tumor growth, metastasis, and angiogenesis in NSCLC. The activation of key signaling pathway under fibronectin was examined by WB assay. RNA-seq was applicated to screening the target gene of fibronectin. Rescue experiment was performed to confirm the role of target gene in fibronectin-mediated function in NSCLC. Finally, luciferase and CHIP assays were used to elucidate the mechanism by which fibronectin regulated the target gene. Results Our results revealed that fibronectin was up-regulated in cancer tissues compared with the normal ones in NSCLC patients. Dish- coated fibronectin enhanced the tumor growth, metastasis, and angiogenesis of NSCLC in vitro and in vivo by promoting EMT and maintaining stemness of NSCLC cells. As expected, fibronectin activated FAK and its downstream MAPK/ERK signaling pathway. WISP3 was screened as a potential target gene of fibronectin. Interestingly, WISP3 effectively activated Wnt signaling pathway, and knockdown of WISP3 effectively blocked the influence of fibronectin on the migration, invasion and vascular structure formation potential of NSCLC cells. Our data also manifested that fibronectin elevated the transcription of WISP3 gene by promoting the binding of HIF-1α to the promoter region of WISP3 in NSCLC cells. Conclusions Our findings sketched the outline of the route for fibronectin exert its role in NSCLC, in which fibronectin activated downstream FAK and MAPK/ERK signaling pathways, and mediated the accumulation of HIF-1α. Then, HIF-1α enabled the transcription of WISP3, and subsequently promoted the activation of Wnt signaling pathway, and finally enhanced the tumor growth, metastasis, and angiogenesis in NSCLC.

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“…Sana et al used RGD and fibronectin-immobilized chitosan scaffolds to piggyback human dental pulp stem cells (hDPSCs) and found the scaffolds can significantly promote cell migration, proliferation, and differentiation [20]. Our team [21] combined FNIII10 and CC (human collagen-like protein) proteins to obtain FCP. The ECM-mimetic FCP provides a microenvironment that supports stem cell localization, proliferation, differentiation, and stemness maintenance.…”

Section: Introductionmentioning

confidence: 99%

A Thermally Stable Recombinant Human Fibronectin Peptide-Fused Protein (rhFN3C) for Faster Aphthous Ulcer (AU) Healing

Cai,

Zhu,

Luo

et al. 2023

Bioengineering

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Approximately 59.4–100% of head and neck cancer patients receiving radiotherapy or radio chemotherapy suffer from aphthous ulcers (AUs), which seriously affect the subsequent treatment. At the same time, AUs are a common oral mucosal disease with a high incidence rate among the population, often accompanied by severe pain, and affect both physical and mental health. Strategies to increase the ulcer healing rate and relieve pain symptoms quickly is a long-term clinical objective. Oral mucosal discontinuity is the main histological hallmark of AUs. So, covering the inner mucosal defect with an in vitro engineered oral mucosal equivalent shows good prospects for AU alleviation. Fibronectin (FN) is a glycopeptide in the extracellular matrix and exhibits opsonic properties, aiding the phagocytosis and clearance of foreign pathogens through all stages of ulcer healing. But native FN comes from animal blood, which has potential health risks. rhFN3C was designed with multi-domains of native FN, whose core functions are the recruitment of cells and growth factors to accelerate AU healing. rhFN3C is a peptide-fused recombinant protein. The peptides are derived from the positions of 1444–1545 (FNIII10) and 1632–1901 (FNIII12–14) in human native FN. We optimized the fermentation conditions of rhFN3C in E. coli BL21 to enable high expression levels. rhFN3C is thermally stable and nontoxic for L929, strongly promotes the migration and adhesion of HaCaT, decreases the incidence of wound infection, and shortens the mean healing time by about 2 days compared to others (p < 0.01). rhFN3C may have great potential for use in the treatment of AUs. The specific methods and mechanisms of rhFN3C are yet to be investigated.

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Recombinant expression a novel fibronectin—collage fusion peptide modulating stem cell stemness via integrin β3

Cited by 7 publications

References 40 publications

Production and Functional Analysis of Collagen Hexapeptide Repeat Sequences in Pichia pastoris

Production and Functional Analysis of Collagen Hexapeptide Repeat Sequences in Pichia pastoris

Fibronectin promotes tumor angiogenesis and progression of non-small-cell lung cancer by elevating WISP3 expression via FAK/MAPK/ HIF-1α axis and activating wnt signaling pathway

A Thermally Stable Recombinant Human Fibronectin Peptide-Fused Protein (rhFN3C) for Faster Aphthous Ulcer (AU) Healing

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