{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,22]],"date-time":"2026-09-22T04:51:34Z","timestamp":1790052694082,"version":"4.0.1"},"reference-count":68,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2023,2,23]],"date-time":"2023-02-23T00:00:00Z","timestamp":1677110400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"The Macular Society"},{"name":"The Humane Research Trust"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Cystatin C, a secreted cysteine protease inhibitor, is abundantly expressed in retinal pigment epithelium (RPE) cells. A mutation in the protein\u2019s leader sequence, corresponding to formation of an alternate variant B protein, has been linked with an increased risk for both age-related macular degeneration (AMD) and Alzheimer\u2019s disease (AD). Variant B cystatin C displays intracellular mistrafficking with partial mitochondrial association. We hypothesized that variant B cystatin C interacts with mitochondrial proteins and impacts mitochondrial function. We sought to determine how the interactome of the disease-related variant B cystatin C differs from that of the wild-type (WT) form. For this purpose, we expressed cystatin C Halo-tag fusion constructs in RPE cells to pull down proteins interacting with either the WT or variant B form, followed by identification and quantification by mass spectrometry. We identified a total of 28 interacting proteins, of which 8 were exclusively pulled down by variant B cystatin C. These included 18 kDa translocator protein (TSPO) and cytochrome B5 type B, both of which are localized to the mitochondrial outer membrane. Variant B cystatin C expression also affected RPE mitochondrial function with increased membrane potential and susceptibility to damage-induced ROS production. The findings help us to understand how variant B cystatin C differs functionally from the WT form and provide leads to RPE processes adversely affected by the variant B genotype.<\/jats:p>","DOI":"10.3390\/cells12050713","type":"journal-article","created":{"date-parts":[[2023,2,24]],"date-time":"2023-02-24T03:01:01Z","timestamp":1677207661000},"page":"713","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Analysis of Wild Type and Variant B Cystatin C Interactome in Retinal Pigment Epithelium Cells Reveals Variant B Interacting Mitochondrial Proteins"],"prefix":"10.3390","volume":"12","author":[{"given":"Emil","family":"Carlsson","sequence":"first","affiliation":[{"name":"Ocular Molecular Biology and Mechanisms of Disease Group, Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool L69 3BX, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Umar","family":"Sharif","sequence":"additional","affiliation":[{"name":"Ocular Molecular Biology and Mechanisms of Disease Group, Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool L69 3BX, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3789-8312","authenticated-orcid":false,"given":"Wasu","family":"Supharattanasitthi","sequence":"additional","affiliation":[{"name":"Ocular Molecular Biology and Mechanisms of Disease Group, Institute of Life Course and Medical Sciences, University of Liverpool, Liverpool L69 3BX, UK"},{"name":"Department of Physiology, Faculty of Pharmacy, Mahidol University, Nakhon Pathom 73170, Thailand"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7568-7116","authenticated-orcid":false,"given":"Luminita","family":"Paraoan","sequence":"additional","affiliation":[{"name":"Ocular Molecular Biology and Mechanisms of Disease Group, Department of Biology, Faculty of Arts and Sciences, Edge Hill University, Ormskirk L39 4QP, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,2,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1186\/s40662-016-0063-5","article-title":"Epidemiology of age-related macular degeneration (AMD): Associations with cardiovascular disease phenotypes and lipid factors","volume":"3","author":"Pennington","year":"2016","journal-title":"Eye Vision"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"26","DOI":"10.1016\/j.neuron.2012.06.018","article-title":"Mechanisms of age-related macular degeneration","volume":"75","author":"Ambati","year":"2012","journal-title":"Neuron"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1111\/j.1755-3768.2011.02179.x","article-title":"Molecular mechanisms of retinal pigment epithelium damage and development of age-related macular degeneration","volume":"90","author":"Kinnunen","year":"2012","journal-title":"Acta Ophthalmol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"151","DOI":"10.1146\/annurev-genom-090413-025610","article-title":"Age-related macular degeneration: Genetics and biology coming together","volume":"15","author":"Fritsche","year":"2014","journal-title":"Annu. 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