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Using a USP7-dependent colorectal cancer cell model, the study applies inducible USP7 knockdown and label-free LC-MS/MS proteomics to quantify USP7-dependent proteins. Co-IP, reduced Ajuba levels after USP7 knockdown, and sensitivity to the USP7 inhibitor FT671 identify Ajuba as a USP7 interaction partner and mediating component, with knockdown of USP7 or Ajuba reducing cell-cell adhesion and altering adherens junction-associated proteins.",{"@graph":69,"@context":122},[70,84,105],{"@type":71,"itemListElement":72},"BreadcrumbList",[73,77,79,82],{"item":74,"name":75,"@type":76,"position":8},"https://docshare.wps.com","Home","ListItem",{"item":78,"name":9,"@type":76,"position":14},"https://docshare.wps.com/document/",{"item":80,"name":35,"@type":76,"position":81},"https://docshare.wps.com/document/healthcare/",3,{"item":83,"name":65,"@type":76,"position":19},"https://docshare.wps.com/document/knockdown-proteomics-reveals-usp7-as-a-regulator-of-cell-cell-adhesion-in-colorectal-cancer-via-ajuba/337201/",{"url":83,"name":65,"@type":85,"image":86,"author":91,"headline":65,"publisher":94,"fileFormat":97,"inLanguage":63,"description":67,"dateModified":98,"datePublished":99,"encodingFormat":97,"isAccessibleForFree":100,"interactionStatistic":101},"DigitalDocument",{"url":87,"@type":88,"width":89,"height":90},"https://docshare.wps.com/thumbnails/knockdown-proteomics-reveals-usp7-as-a-regulator-of-cell-cell-adhesion-in-colorectal-cancer-via-ajuba/337201.png","ImageObject",300,407,{"name":92,"@type":93},"Theodora","Person",{"url":74,"name":95,"@type":96},"DocShare","Organization","application/pdf","2026-10-27","2026-09-21",true,{"@type":102,"interactionType":103,"userInteractionCount":24},"InteractionCounter",{"@type":104},"ViewAction",{"@type":106,"mainEntity":107},"FAQPage",[108,114,118],{"name":109,"@type":110,"acceptedAnswer":111},"What experimental approach was used to identify USP7-dependent proteins?","Question",{"text":112,"@type":113},"The study used inducible USP7 knockdown coupled with LC-MS/MS to quantify proteins whose abundance depends on USP7 activity.","Answer",{"name":115,"@type":110,"acceptedAnswer":116},"How was Ajuba determined to interact with USP7?",{"text":117,"@type":113},"Ajuba was identified as a USP7 interaction partner using co-IP, and its protein levels dropped substantially after USP7 knockdown. Ajuba also showed sensitivity to the specific USP7 inhibitor FT671.",{"name":119,"@type":110,"acceptedAnswer":120},"What effect does USP7 knockdown have on colorectal cancer cells?",{"text":121,"@type":113},"USP7 knockdown significantly inhibited cell viability and colony formation and reduced cell-cell adhesion, alongside changes in other proteins associated with adherens junctions.","https://schema.org",{"og:url":83,"og:type":124,"og:title":65,"og:site_name":95,"og:description":67},"article",{"robots":126,"canonical":83},"index,follow",{"doc_id":128,"site_id":62},337201,1791570359,{"code":4,"msg":5,"data":131},{"doc_id":128,"user_id":132,"nickname":92,"user_avatar":133,"doc_module":4,"category_id":34,"category_name":35,"doc_title":65,"doc_description":67,"doc_content":134,"file_id":135,"file_url":136,"file_type":137,"file_size":138,"view_count":24,"is_deleted":4,"is_public":8,"is_downloadable":8,"audit_status":8,"page_count":139,"language":140,"language_code":63,"site_id":62,"html_lang":63,"table_of_contents":141,"faqs":142,"seo_title":143,"seo_description":67,"update_tm":144,"read_time":145},687197207919,"https://ap-avatar.wpscdn.com/avatar/a000253d6f5f7c60be?x-image-process=image/resize,m_fixed,w_180,h_180&k=1779446848396160552","Knockdown Proteomics Reveals USP7 as a Regulator of Cell-Cell Adhesion in Colorectal Cancer via AJUBA  \nAuthors  \nAhood Al-Eidan, Ben Draper, Siyuan Wang, Brandon Coke, Paul Skipp, Yihua Wang, and Rob M. Ewing  \nCorrespondence  \n[rob.ewing@soton.ac.uk](rob.ewing@soton.ac.uk)  \nIn Brief  \nThe deubiquitinase, USP7, regulates the abundance of cancer-associated proteins and is a key target in multiple types of tumor. Using USP7 knockdown coupled to label-free proteomicsin a USP7-dependent cancer colorectal cell-line, new interaction partners of USP7 can be identified. These include Ajuba, a cell-cell adhesion scaffold protein identified as a USP7 interaction partner, thereby mediating the regulation of cell-cell adhesion by USP7 .  \nGraphical Abstract  \nHighlights  \n• The deubiquitinase, USP7, is a key cancer target in many solid tumors.  \n• Knockdown proteomics can reveal novel USP7 interaction partners.  \n• USP7 interacts with Ajuba and thereby regulates cell-cell adhesion.  \n2024, Mol Cell Proteomics 23(12), 100878  \n© 2024 THE AUTHORS. Published by Elsevier Inc on behalf of American Society for Biochemistry and Molecular Biology. This is an open access article under the CC BY-NC-ND license ([http://](http://)[ ](http://)[creativecommons.org/licenses/by-nc-nd/4.0/](creativecommons.org/licenses/by-nc-nd/4.0/)).  \n[https://doi.org/10.1016/j.mcpro.2024.100878](https://doi.org/10.1016/j.mcpro.2024.100878)  \n RESEARCH  \n| Knockdown Proteomics Reveals USP7 as a Regulator of Cell-Cell Adhesion in Colorectal Cancer via AJUBA\u003Cbr>Ahood Al-Eidan 1 ,2 , Ben Draper 1, Siyuan Wang 1 , Brandon Coke 1 , Paul Skipp 1 , Yihua Wang 1, and Rob M. Ewing 1 ,* |  |\n| --- | --- |\n| Ubiquitin-specific protease 7 (USP7) is implicated in many cancers including colorectal cancer in which it regulates cellular pathways such as Wnt signaling and the P53– MDM2 pathway. With the discovery of small-molecule inhibitors, USP7 has also become a promising target for cancer therapy and therefore systematically identifying USP7 deubiquitinase interaction partners and substrates has become an important goal. In this study, we selected a colorectal cancer cell model that is highly dependent on USP7 and in which USP7 knockdown significantly inhibited colorectal cancer cell viability, colony formation, and cell-cell adhesion. We then used inducible knockdown of USP7 followed by LC-MS/MS to quantify USP7-dependent proteins. We identified the Ajuba LIM domain protein as an interacting partner of USP7 through co-IP, its substantially reduced protein levels in response to USP7 knockdown, and its sensitivity to the specific USP7 inhibitor FT671. The Ajuba protein has been shown to have oncogenic functions in colorectal and other tumors, including regulation of cell-cell adhesion. We show that both knockdown of USP7 or Ajuba results in a substantial reduction of cell-cell adhesion, with concomitant effects on other proteins associated with adherens junctions. Our findings underlie the role of USP7 in colorectal cancer through its protein interaction networks and show that the Ajuba protein is a component of USP7 protein networks present in colorectal cancer.\u003Cbr>There has been rapidly increasing interest in the role of deubiquitinating enzymes (DUBs) in many types of tumor including colorectal cancer (CRC) (1) . DUB proteins are proteases that reverse E3 ligase activity by cleaving a single ubiquitin, polyubiquitin chains or ubiquitin-like modifications on target proteins (2–4) . This role of DUBs is similar to the regulatory role of the phosphatases in a kinase/phosphatase pathway (3). There are nearly 100 DUBs in the human genome, grouped into five families: ubiquitin-specific protease (USP), OUT, UCH, Josephin (cysteine proteases), and JAMM (a family of metalloproteases); DUBs have high specificity for | ubiquitin. Different DUBs have different domains for the protein–protein interactions, substrate specificity, and cellular localization. Quantitative studies of multiple aspects of DUB","cbCaieVKvEbKLXbW","https://ap.wps.com/l/cbCaieVKvEbKLXbW","pdf",3109881,17,"English","# Research\n## USP7 role in colorectal cancer\n## Experimental strategy: inducible knockdown and proteomics\n## Identification and validation of Ajuba\n## Mechanistic implications for adhesion networks","[{\"question\":\"What experimental approach was used to identify USP7-dependent proteins?\",\"answer\":\"The study used inducible USP7 knockdown coupled with LC-MS/MS to quantify proteins whose abundance depends on USP7 activity.\"},{\"question\":\"How was Ajuba determined to interact with USP7?\",\"answer\":\"Ajuba was identified as a USP7 interaction partner using co-IP, and its protein levels dropped substantially after USP7 knockdown. Ajuba also showed sensitivity to the specific USP7 inhibitor FT671.\"},{\"question\":\"What effect does USP7 knockdown have on colorectal cancer cells?\",\"answer\":\"USP7 knockdown significantly inhibited cell viability and colony formation and reduced cell-cell adhesion, alongside changes in other proteins associated with adherens junctions.\"}]","Knockdown Proteomics Reveals USP7 as a Regulator of Cell-Cell Adhesion in Colorectal Cancer via AJUBA | PDF",1790018113,43]