{"doi":"10.1101/2020.02.07.935932","title":"Probing the effect of glycosaminoglycan depletion on integrin interactions with collagen I fibrils in the native ECM environment","abstract":"Abstract Fibrillar collagen–integrin interactions in the extracellular matrix (ECM) regulate a multitude of cellular processes and cell signalling. Collagen I fibrils serve as the molecular scaffolding for connective tissues throughout the human body and are the most abundant protein building blocks in the ECM. The ECM environment is diverse, made up of several ECM proteins, enzymes, and proteoglycans. The contents of the ECM environment are modulated by disease and aging and may influence these critical collagen–integrin interactions. In particular, glycosaminoglycans (GAGs), anionic polysaccharides that decorate proteoglycans, become depleted in the ECM with natural aging and their mis-regulation has been linked to cancers and other diseases. The impact of GAG concentration in the ECM environment on collagen interactions is not well understood. Here, we integrate protein adhesion assays with liquid high resolution atomic force microscopy (AFM) to assess the affects of GAG depletion on the interaction of collagen I fibrils with the integrin α 2 I domain. Adhesion assays demonstrate that α 2 I preferentially binds to GAG-depleted collagen I fibrils. By amplitude modulated AFM in air and in solution, we find that GAG-depleted collagen I fibrils retain structural features of the native fibrils, including their characteristic D-banding pattern, a key structural motif. AFM fast force mapping in solution shows that GAG depletion reduces the stiffness of individual fibrils, lowering the indentation modulus by half compared to native fibrils. Together these results shed new light on how GAGs influence collagen I fibril– integrin interactions and may aid in strategies to treat diseases that result from GAG misregulation. Statement for broader audience Aging and disease result in mis-regulation of glycosaminoglycan (GAG) levels in the extracellular matrix (ECM), which may affect fibrillar collagen interactions that are vital for cellular processes. Here, we characterize the impact of GAG depletion on collagen–integrin α 2 I domain interactions and collagen fibril topography and stiffness. We show that GAG depletion increases collagen–α 2 I binding and reduces stiffness in comparison to native fibrils. These results may inform on strategies for treating GAG mis-regulation.","journal":"bioRxiv (Cold Spring Harbor Laboratory)","year":2020,"id":128860,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":0,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9668,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2020-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":579587,"name":"Cody L. Hoop","orcid":"0000-0003-4441-8321","position":1,"is_corresponding":false},{"id":553460,"name":"Jonathan K. Williams","orcid":"0000-0002-7272-6885","position":2,"is_corresponding":false},{"id":580117,"name":"Robert J. Hayes","orcid":null,"position":3,"is_corresponding":false},{"id":332821,"name":"Jean Baum","orcid":null,"position":4,"is_corresponding":false},{"id":579586,"name":"Jonathan D. Roth","orcid":"0000-0002-1983-385X","position":0,"is_corresponding":true}],"reference_count":74,"raw_metadata":null,"created_at":"2026-07-18T23:15:42.522809Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}