title: The C-terminus hot spot region helps in the fibril formation of bacteriophage-associated hyaluronate lyase (HylP2).
abstract: 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 ((N)HylP2(C)HylP and (N)HylP(C)HylP2) or HylP did not form fibrils at acidic pH. However, seeding of prefibrils of HylP2 recompensed nucleation and led to fibrillation in (N)HylP(C)HylP2. 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.
authors.affiliation: Molecular and Structural Biology Division, CSIR-Central Drug Research Institute, Sector 10, Jankipuram Extension, Lucknow, INDIA, PIN 226 03.
authors.forename: Harish
authors.initials: H
authors.lastname: Shukla
authors.affiliation: Molecular and Structural Biology Division, CSIR-Central Drug Research Institute, Sector 10, Jankipuram Extension, Lucknow, INDIA, PIN 226 03.
authors.forename: Sudhir Kumar
authors.initials: SK
authors.lastname: Singh
authors.affiliation: Molecular and Structural Biology Division, CSIR-Central Drug Research Institute, Sector 10, Jankipuram Extension, Lucknow, INDIA, PIN 226 03.
authors.forename: Amit Kumar
authors.initials: AK
authors.lastname: Singh
authors.affiliation: Sophisticated Analytical Instrument Facility , CSIR-Central Drug Research Institute, Sector 10, Jankipuram Extension, Lucknow, INDIA, PIN 226 031.|Academy of Scientific and Innovative Research, CSIR-Central Drug Research Institute, Sector 10, Jankipuram Extension, Lucknow, INDIA, PIN 226 031.
authors.forename: Kalyan
authors.initials: K
authors.lastname: Mitra
authors.affiliation: Molecular and Structural Biology Division, CSIR-Central Drug Research Institute, Sector 10, Jankipuram Extension, Lucknow, INDIA, PIN 226 03.|Academy of Scientific and Innovative Research, CSIR-Central Drug Research Institute, Sector 10, Jankipuram Extension, Lucknow, INDIA, PIN 226 031.
authors.forename: Md Sohail
authors.initials: MS
authors.lastname: Akhtar
mesh_terms: D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000201:enzymology* / Q000235:genetics*; D001483:Base Sequence; D006025:Glycosaminoglycans / Q000378:metabolism; D006820:Hyaluronic Acid / Q000378:metabolism; D009154:Mutation; D011133:Polysaccharide-Lyases / Q000235:genetics / Q000378:metabolism*; D066329:Protein Aggregates / Q000502:physiology*; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D013297:Streptococcus pyogenes / Q000821:virology
chemical_list: D006025:Glycosaminoglycans; D066329:Protein Aggregates; D006820:Hyaluronic Acid; D011133:Polysaccharide-Lyases; C100002:hyaluronate lyase