{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,4]],"date-time":"2026-06-04T01:00:29Z","timestamp":1780534829154,"version":"3.54.1"},"reference-count":36,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2025,8,29]],"date-time":"2025-08-29T00:00:00Z","timestamp":1756425600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Foods"],"abstract":"<jats:p>The epidermal growth factor receptor (EGFR), as an important target protein for inhibiting and intervening in osteoporosis, is associated with cell migration, proliferation, and apoptosis. Peptides derived from food have been shown to have a strong affinity for EGFR, thereby regulating downstream cellular-signaling pathways and participating in stimulating bone formation. However, it is still a \u201cblack box\u201d as to how active peptides affect the conformational changes in the EGFR-binding domain when interacting with its ligand EGF. To shed light on the roles, peptides in EGFR binding, which is involved in the osteoblast differentiation, a high EGFR affinity soybean peptide (HEP) was isolated and purified from soy yogurt. Firstly, the osteogenic activity of HEP was identified through cellular alkaline-phosphatase (ALP) and calcium influx. HEP promoted ALP activity from 0.01897 \u00b1 0.00165 to 0.04051 \u00b1 0.00402 U\/mg after 100 \u03bcM of peptide treatment, and free intracellular calcium ions and calcium deposition both increased in a dose-dependent manner at 1\u2013100 \u03bcg\/mL. Secondly, the interaction between HEP and EGFR was detected by bioinformatics, spectroscopy analysis, and Western blot. The Molecular docking results showed that HEP (VVELLKAFEEKF) exhibited high affinity among all the peptides, with -CDOCKER energy values of 184.077 kcal\/mol on one EGFR. Moreover, a different loop conformation has been detected in HEP, comparing it to that of EGF, which influences HEP interactions with EGFR. GlU3, LEU4, and LEU5 (HEP) match GLU40, LEU26, and GLU40 (EGF). Moreover, the CD data showed that HEP could interact with extracellular domain protein of EGFR, but the secondary structure did not change after HEP was mixed with Mutant extracellular domain protein. Furthermore, treatment with HEP increased the expression of EGFR and the activation of the PI3K-RUNX2-signaling pathway. These results suggested that HEP may have the function of promoting bone remodeling, which could promote the binding between EGF and EGFR and may be used as a potential active factor for functional food development to prevent osteoporosis.<\/jats:p>","DOI":"10.3390\/foods14173032","type":"journal-article","created":{"date-parts":[[2025,8,29]],"date-time":"2025-08-29T09:08:52Z","timestamp":1756458532000},"page":"3032","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Interactions Between HEP Peptide and EGFR Involved in the Osteoblast Differentiation"],"prefix":"10.3390","volume":"14","author":[{"given":"Jing","family":"Gan","sequence":"first","affiliation":[{"name":"Heilongjiang Green Food Science Research Institute, Harbin 150028, China"},{"name":"College of Life Science, Yantai University, Yantai 264000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8805-9351","authenticated-orcid":false,"given":"Yanling","family":"Huang","sequence":"additional","affiliation":[{"name":"Heilongjiang Green Food Science Research Institute, Harbin 150028, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mengqi","family":"Jian","sequence":"additional","affiliation":[{"name":"College of Life Science, Yantai University, Yantai 264000, China"},{"name":"Beijing Key Laboratory of Functional Food from Plant Resources, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuhang","family":"Chen","sequence":"additional","affiliation":[{"name":"College of Life Science, Yantai University, Yantai 264000, China"},{"name":"Beijing Key Laboratory of Functional Food from Plant Resources, College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuxuan","family":"Jiang","sequence":"additional","affiliation":[{"name":"College of Life Science, Yantai University, Yantai 264000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Qiao","sequence":"additional","affiliation":[{"name":"College of Life Science, Yantai University, Yantai 264000, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yang","family":"Li","sequence":"additional","affiliation":[{"name":"Heilongjiang Green Food Science Research Institute, Harbin 150028, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2025,8,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1576","DOI":"10.1016\/j.stemcr.2022.06.001","article-title":"The role of CDK8 in mesenchymal stem cells in controlling osteoclastogenesis and bone homeostasis","volume":"17","author":"Yamada","year":"2022","journal-title":"Stem Cell Rep."},{"key":"ref_2","doi-asserted-by":"crossref","unstructured":"Gao, Y., Patil, S., and Jia, J. (2021). The Development of Molecular Biology of Osteoporosis. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22158182"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1016\/j.ijbiomac.2017.02.056","article-title":"Pilose antler peptide protects osteoblasts from inflammatory and oxidative injury through EGF\/EGFR signaling","volume":"99","author":"Chunhui","year":"2017","journal-title":"Int. J. Biol. Macromol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"11137","DOI":"10.1096\/fj.201900436RR","article-title":"The role of EGFR signaling in age-related osteoporosis in mouse cortical bone","volume":"33","author":"Liu","year":"2019","journal-title":"FASEB J."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2288","DOI":"10.1074\/jbc.M117.811299","article-title":"Ligand-activated epidermal growth factor receptor (EGFR) signaling governs endocytic trafficking of unliganded receptor monomers by non-canonical phosphorylation","volume":"293","author":"Tanaka","year":"2018","journal-title":"J. Biol. Chem."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2414","DOI":"10.1111\/iwj.14075","article-title":"The use of epidermal growth factor in dermatological practice","volume":"20","author":"Shin","year":"2023","journal-title":"Int. Wound J."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"557","DOI":"10.1016\/j.cell.2012.12.030","article-title":"Architecture and Membrane Interactions of the EGF Receptor","volume":"152","author":"Arkhipov","year":"2013","journal-title":"Cell"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Rybak, J.A., Sahoo, A.R., Kim, S., Pyron, R.J., Pitts, S.B., Guleryuz, S., Smith, A.W., Buck, M., and Barrera, F.N. (2023). Allosteric inhibition of the epidermal growth factor receptor through disruption of transmembrane interactions. J. Biol. Chem., 299.","DOI":"10.1016\/j.jbc.2023.104914"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"815","DOI":"10.2174\/1568026620666200303123102","article-title":"Review on Epidermal Growth Factor Receptor (EGFR) Structure, Signaling Pathways, Interactions, and Recent Updates of EGFR Inhibitors","volume":"20","author":"Sabbah","year":"2020","journal-title":"Curr. Top. Med. Chem."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2127","DOI":"10.1016\/j.bpj.2020.10.008","article-title":"Autophosphorylation of EGFR at Y954 Facilitated Homodimerization and Enhanced Downstream Signals","volume":"119","author":"Thirukkumaran","year":"2020","journal-title":"Biophys. J."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1721","DOI":"10.3892\/ijo.2015.2871","article-title":"EGF-stimulated AKT activation is mediated by EGFR recycling via an early endocytic pathway in a gefitinib-resistant human lung cancer cell line","volume":"46","author":"Nishimura","year":"2015","journal-title":"Int. J. Oncol."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"24462","DOI":"10.1021\/acsami.7b05617","article-title":"EGFR\/EGFRvIII Dual-Targeting Peptide-Mediated Drug Delivery for Enhanced Glioma Therapy","volume":"9","author":"Mao","year":"2017","journal-title":"ACS Appl. Mater. Interfaces"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"7179","DOI":"10.1021\/acs.jafc.7b02067","article-title":"Isolation and Characterization of Lactoferrin Peptides with Stimulatory Effect on Osteoblast Proliferation","volume":"65","author":"Fan","year":"2017","journal-title":"J. Agric. Food Chem."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"e1900525","DOI":"10.1002\/mnfr.201900525","article-title":"Duck Egg White-Derived Peptide VSEE (Val-Ser-Glu-Glu) Regulates Bone and Lipid Metabolisms by Wnt\/\u03b2-Catenin Signaling Pathway and Intestinal Microbiota","volume":"63","author":"Guo","year":"2019","journal-title":"Mol. Nutr. Food Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"15611","DOI":"10.1021\/acs.jafc.1c06028","article-title":"Characterizations and the Mechanism Underlying Osteogenic Activity of Peptides from Enzymatic Hydrolysates of Stichopus japonicus","volume":"69","author":"Yang","year":"2021","journal-title":"J. Agric. Food Chem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"7630","DOI":"10.1021\/acs.jafc.0c02717","article-title":"Cell Proliferation Stimulation Ability and Osteogenic Activity of Low Molecular Weight Peptides Derived from Bovine Gelatin Hydrolysates","volume":"68","author":"Wang","year":"2020","journal-title":"J. Agric. Food Chem."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Wang, K., Kong, X., Du, M., Yu, W., Wang, Z., Xu, B., Yang, J., Xu, J., Liu, Z., and Cheng, Y. (2022). Novel Soy Peptide CBP: Stimulation of Osteoblast Differentiation via T\u03b2RI-p38-MAPK-Depending RUNX2 Activation. Nutrients, 14.","DOI":"10.3390\/nu14091940"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"23246","DOI":"10.1021\/acs.jafc.4c04882","article-title":"Soy Peptide Ameliorate TGF-\u03b21-Mediated Osteoblast Differentiation through Smad and MAPK Signaling Pathways","volume":"72","author":"Wang","year":"2024","journal-title":"J. Agric. Food Chem."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"2939","DOI":"10.1002\/jsfa.12370","article-title":"Isolation, characterization, and molecular docking analyses of novel calcium-chelating peptide from soy yogurt and the study of its calcium chelation mechanism","volume":"103","author":"Gan","year":"2023","journal-title":"J. Sci. Food Agric."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"32075","DOI":"10.1074\/jbc.M110.130435","article-title":"Role of H-1 and H-2 subunits of soybean seed ferritin in oxidative deposition of iron in protein","volume":"285","author":"Deng","year":"2010","journal-title":"J. Biol. Chem."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"N\u2019Deh, K.P.U., Yoo, H.S., Chung, K.H., Lee, K.J., Kim, D.H., Yoon, J.A., and An, J.H. (2020). Collagen Extract Derived from Yeonsan Ogye Chicken Increases Bone Microarchitecture by Suppressing the RANKL\/OPG Ratio via the JNK Signaling Pathway. Nutrients, 12.","DOI":"10.3390\/nu12071967"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Kong, X., Xiao, Z., Chen, Y., Du, M., Zhang, Z., Wang, Z., Xu, B., Cheng, Y., Yu, T., and Gan, J. (2023). Calcium-binding properties, stability, and osteogenic ability of phosphorylated soy peptide-calcium chelate. Front. Nutr., 10.","DOI":"10.3389\/fnut.2023.1129548"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1038\/s41421-024-00689-6","article-title":"Cell signaling and transcriptional regulation of osteoblast lineage commitment, differentiation, bone formation, and homeostasis","volume":"10","author":"Zhu","year":"2024","journal-title":"Cell Discov."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Xu, Z., Han, S., Chen, H., Zhu, Z., Han, L., Dong, X., Du, M., and Li, T. (2022). Characterization of Chelation and Absorption of Calcium by a Mytilus edulis Derived Osteogenic Peptide. Front. Nutr., 9.","DOI":"10.3389\/fnut.2022.840638"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1349","DOI":"10.1002\/jsfa.10745","article-title":"Xanthine oxidase inhibitory peptides derived from tuna protein: Virtual screening, inhibitory activity, and molecular mechanisms","volume":"101","author":"Yu","year":"2021","journal-title":"J. Sci. Food Agric."},{"key":"ref_26","doi-asserted-by":"crossref","unstructured":"Khaleghiabbasabadi, M., Taghavian, H., Gholami, P., Khodabakhshi, S., Gheibi, M., Wac\u0142awek, S., \u010cern\u00edk, M., Silvestri, D., Raczak, K.B., and Moezzi, R. (2024). A Novel Organic-Inorganic-Nanocomposite-Based Reduced Graphene Oxide as an Efficient Nanosensor for NO(2) Detection. Nanomaterials, 14.","DOI":"10.3390\/nano14241983"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Wang, Y., Wang, Y., Zhuang, X., Zhang, Y., Fang, B., and Fu, Y. (2025). Unraveling the Osteogenic Activity and Molecular Mechanism of an Antioxidant Collagen Peptide in MC3T3-E1 Cells. Nutrients, 17.","DOI":"10.3390\/nu17050824"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"834","DOI":"10.1111\/1750-3841.15067","article-title":"Desalted duck egg white peptides promoted osteogenesis via wnt\/\u03b2-catenin signal pathway","volume":"85","author":"Guo","year":"2020","journal-title":"J. Food Sci."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"121711","DOI":"10.1016\/j.carbpol.2023.121711","article-title":"Impact of high temperature on microstructural changes and oil absorption of tigernut (Cyperus esculentus L.) starch: Investigations in the starch-oil model system","volume":"328","author":"Wu","year":"2024","journal-title":"Carbohydr. Polym."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1240","DOI":"10.2174\/1570180820666230224100942","article-title":"Molecular Docking, Molecular Dynamics Simulation, and Analysis of EGFR-derived Peptides against the EGF","volume":"21","author":"Samaneh","year":"2024","journal-title":"Lett. Drug Des. Discov."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"145011","DOI":"10.1016\/j.foodchem.2025.145011","article-title":"ACE-inhibitory peptides from Morchella esculenta: Screening, kinetics, and molecular dynamics simulation","volume":"490","author":"Zeng","year":"2025","journal-title":"Food Chem."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1007\/s11010-014-2210-4","article-title":"EGFR-AKT-mTOR activation mediates epiregulin-induced pleiotropic functions in cultured osteoblasts","volume":"398","author":"Fan","year":"2015","journal-title":"Mol. Cell. Biochem."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Zhao, H., Jia, P., Nanding, K., Wu, M., Bai, X., Morigen, M., and Fan, L. (2022). Lysophosphatidic acid suppresses apoptosis of high-grade serous ovarian cancer cells by inducing autophagy activity and promotes cell-cycle progression via EGFR-PI3K\/Aurora-A(Thr288)-geminin dual signaling pathways. Front. Pharmacol., 13.","DOI":"10.3389\/fphar.2022.1046269"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"640","DOI":"10.3109\/10799893.2015.1041647","article-title":"The PI3K\/AKT cell signaling pathway is involved in regulation of osteoporosis","volume":"35","author":"Xi","year":"2015","journal-title":"J. Recept. Signal Transduct."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"169","DOI":"10.1007\/s00223-006-0083-6","article-title":"Runx2 overexpression enhances osteoblastic differentiation and mineralization in adipose--derived stem cells in vitro and in vivo","volume":"79","author":"Zhang","year":"2006","journal-title":"Calcif. Tissue Int."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"71","DOI":"10.1002\/jbmr.2321","article-title":"Loss of Runx2 in committed osteoblasts impairs postnatal skeletogenesis","volume":"30","author":"Adhami","year":"2015","journal-title":"J. Bone Miner. Res."}],"container-title":["Foods"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2304-8158\/14\/17\/3032\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:35:01Z","timestamp":1760034901000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2304-8158\/14\/17\/3032"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,8,29]]},"references-count":36,"journal-issue":{"issue":"17","published-online":{"date-parts":[[2025,9]]}},"alternative-id":["foods14173032"],"URL":"https:\/\/doi.org\/10.3390\/foods14173032","relation":{},"ISSN":["2304-8158"],"issn-type":[{"value":"2304-8158","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,8,29]]}}}