Dynamic light scattering study of the two-domain structure of Humicola insolens endoglucanase V

• Published in: FEBS letters Vol. 376; no. 1; pp. 49 - 52
• Main Authors: Boisset, Claire, Borsali, Redouane, Schülein, Martin, Henrissat, Bernard
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 27.11.1995; Wiley
• Subjects: Binding Sites; Cellulase; Cellulase - chemistry; Cloning, Molecular; Domain structure; Dynamic light scattering; Endoglucanase; Fungal Proteins - chemistry; Fungi - enzymology; Gene Expression Regulation, Fungal - genetics; Humicola insolens; Recombinant Proteins - genetics; Scattering, Radiation; Endoglucanase; Domain structure; Dynamic light scattering; Humicola insolens; Cellulase
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/0014-5793(95)01244-0

Endoglucanase V (EG V) of Humicola insolens is composed of a catalytic domain and of a cellulose-binding domain linked by a 33 amino acid long peptide rich in Ser, Thr and Pro residues. This work describes the dynamic behavior of the two-domain structure of EG V as revealed by quasi-elastic light scattering experiments. For both the full-length and the isolated catalytic domain, the autocorrelation function is essentially described by a single relaxation mode. The equivalent hydrodynamic radius of the catalytic domain was found to correspond precisely to the dimensions measured from the previously determined three-dimensional structure. The results obtained with the full-length protein allow a description of the two domain structure of EG V similar to that resulting from earlier studies using small angle X-ray scattering on cellulases from Trichoderma reesei. The hydrodynamic dimensions of the entire enzyme can be approximated as an ellipsoid with dimensions of 42 × 133.6 A ̊ .