文章基本信息

标题：The C-terminus hot spot region helps in the fibril formation of bacteriophage-associated hyaluronate lyase (HylP2)
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作者：Harish Shukla ; Sudhir Kumar Singh ; Amit Kumar Singh 等
期刊名称：Scientific Reports
电子版ISSN：2045-2322
出版年度：2015
卷号：5
期号：1
DOI：10.1038/srep14429
语种：English
出版社：Springer Nature
摘要：The bacteriophage encoded hyaluronate lyases (HylP and HylP2) degrade hyaluronan and other glycosaminoglycans. HylP2 forms a functional fibril under acidic conditions in which its N-terminus is proposed to form the fibrillar core, leading to nucleation and acceleration of fibril formation. Here we report the presence of a hot spot region (A₁₄₄GVVVY₁₄₉) towards the carboxy terminus of HylP2, essential for the acceleration of fibril formation. The ‘hot spot’ is observed to be inherently mutated for valines (A₁₇₈AMVMY₁₈₃) in case of HylP. The N- terminal swapped chimeras between these phage HLs (^NHylP₂^CHylP and ^NHylP^CHylP2) or HylP did not form fibrils at acidic pH. However, seeding of prefibrils of HylP2 recompensed nucleation and led to fibrillation in ^NHylP^CHylP2. The V147A mutation in the ‘hot spot’ region abolished fibril formation in HylP2. The M179V and M181V double mutations in the ‘hot spot’ region of HylP led to fibrillation with the seeding of prefibrils. It appears that fibrillation in HylP2 even though is initiated by the N-terminus, is accelerated by the conserved ‘hot spot’ region in the C-terminus. A collagenous (Gly-X-Y)₁₀ motif in the N-terminus and a mutated ‘hot spot’ region in the C-terminus of HylP affect fibrillar nucleation and acceleration respectively.