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Title: Unraveling the differential structural stability and dynamics features of T7 endolysin partially folded conformations
Authors: Sharma M.
Kumar D.
Poluri, Krishna Mohan
Published in: Biochimica et Biophysica Acta - General Subjects
Abstract: Background: Characterization of partially collapsed protein conformations at atomic level is a daunting task due to their inherent flexibility and conformational heterogeneity. T7 bacteriophage endolysin (T7L) is a single-domain amidase that facilitates the lysis of Gram-negative bacteria. T7L exhibits a pH-dependent structural transition from native state to partially folded (PF) conformation. In the pH range 5–3, T7L PF states display differential ANS binding characteristics. Methods: CD, fluorescence, NMR spectroscopy and lysis assays were used to investigate the structure-stability- dynamics relationships of T7L PF conformations. Results: Structural studies indicated a partial loss of secondary/tertiary structures compared to its native state. The loss in the tertiary structure and the hydrophobic core opening increases upon decrease of pH from 5 to 3. Thermal denaturation experiments delineated that the pH 5 conformation is thermally irreversible in contrast to pH 3, depicting that hydrophobic core opening is essential for thermal reversibility. Further, urea dependent unfolding features of PF state at pH 5 and 4 evidenced for a collapsed conformation at intermediate urea concentrations. Residue level studies revealed that ?1-helix and ?3-?4 segment of T7L are the major contributors for such a structural collapse and inherent dynamics. Conclusions: The results suggested that the low pH PF states of T7L are heterogeneous and exhibits differential structural, unfolding, thermal reversibility, and dynamic features. General significance: Unraveling the structure-stability characteristics of different endolysin conformations is essential for designing novel chimeric and engineered phage endolysins as broadband antimicrobial agents over a varied pH range. © 2018 Elsevier B.V.
Citation: Biochimica et Biophysica Acta - General Subjects(2018), 1862(4): 924-935
Issue Date: 2018
Publisher: Elsevier B.V.
Keywords: Amidase
Molten globule
Partially folded conformation
T7 endolysin
ISSN: 3044165
Author Scopus IDs: 36840988200
Author Affiliations: Sharma, M., Department of Biotechnology, Indian Institute of Technology Roorkee, Roorkee, Uttarakhand 247667, India
Kumar, D., Centre of Biomedical Research, SGPGIMS Campus, Lucknow, Uttar Pradesh 226014, India
Poluri, K.M., Department of Biotechnol
Funding Details: This work is supported by the SERB - SB/YS/LS-380/2013 , DBT-IYBA fellowship – BT/07/IYBA/2013-19 , and startup aid from MHRD-IITR, Government of India (GoI) to KMP. MS acknowledge the receipt of JRF/SRF fellowship from UGC-NET. We acknowledge the support
Corresponding Author: Poluri, K.M.; Department of Biotechnology, Indian Institute of Technology Roorkee (IIT-Roorkee)India; email:
Appears in Collections:Journal Publications [BT]

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