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Emerging anti\u2010amyloid immunotherapies rely on recruiting immune proteases to clear these pathological aggregates, yet a stark disconnect remains between the high proteolytic efficiency observed in dilute in vitro assays and the limited efficacy of these treatments in vivo. This discrepancy likely arises because standard models neglect the dense macromolecular crowding characteristic of the cellular milieu. Here, we address this gap by investigating the degradation of alpha\u2010synuclein amyloids by key immune proteases (cathepsin B, cathepsin D, and matrix metalloproteinase\u20109) in the presence of synthetic crowding agents at physiological densities. We found that macromolecular crowding in a crowding agent\u2010 and dose\u2010dependent manner inhibits protease\u2010induced amyloid declusterization, cluster size reduction, fibril fragmentation, and structural disorganization. We propose that the observed inhibition arises from the excluded volume effect, which stabilizes fibril structure and restricts both enzyme mobility and steric access to the fibril surface rather than suppressing catalytic activity. By restricting the extent of enzymatic amyloid degradation, this \u201ccrowding shield\u201d modulated the cytotoxic consequences of enzymatic processing in human cell lines, attenuating enzyme\u2010induced shifts in fibril toxicity in a condition\u2010dependent manner. These findings identify the crowded cellular environment as a critical stabilizer of pathogenic protein aggregates, explaining their persistence in vivo.<\/jats:p>","DOI":"10.1002\/pro.70707","type":"journal-article","created":{"date-parts":[[2026,6,23]],"date-time":"2026-06-23T12:32:54Z","timestamp":1782217974000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Macromolecular crowding inhibits degradation of alpha\u2010synuclein amyloid fibrils induced by cathepsins and\n                    <scp>MMP9<\/scp>"],"prefix":"10.1002","volume":"35","author":[{"given":"Maksim I.","family":"Sulatsky","sequence":"first","affiliation":[{"name":"Laboratory of Cell Morphology Institute of Cytology Russian Academy of Sciences  St. Petersburg Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Olesya V.","family":"Stepanenko","sequence":"additional","affiliation":[{"name":"Laboratory of Structural Dynamics, 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