Detergent induces the formation of IgG aggregates: A multi-methodological approach

被引:9
|
作者
Amani, Samreen [1 ]
Nasim, Faisal [1 ]
Khan, Taqi Ahmed [1 ]
Fazili, Naveed Ahmad [1 ]
Furkan, Mohammad [1 ]
Bhat, Imtiyaz Ahmad [3 ]
Khan, Javed Masood [2 ]
Khan, Rizwan Hasan [2 ]
Naeem, Aabgeena [1 ]
机构
[1] Aligarh Muslim Univ, Fac Life Sci, Dept Biochem, Aligarh 202002, Uttar Pradesh, India
[2] Aligarh Muslim Univ, Interdisciplincily Biotechnol Unit, Aligarh 202002, Uttar Pradesh, India
[3] Aligarh Muslim Univ, Fac Sci, Dept Chem, Aligarh 202002, Uttar Pradesh, India
关键词
Aggregation; CHAPSO; Dynamic light scattering; Molten globule; THIOFLAVIN-T; PROTOFIBRILS; PROTEINS; PROTOCOL; FIBRILS; PATHWAY; BINDING; RED;
D O I
10.1016/j.saa.2013.09.141
中图分类号
O433 [光谱学];
学科分类号
0703 ; 070302 ;
摘要
Role of micellar environment created by Triton X-100 (TX-100) and CHAPSO on protein conformation using IgG as a model system has been studied in this paper. A substantial amount of secondary structure with the reduction in constant tertiary contacts was obtained in both bovine and human IgG in the presence of 0.12 mM TX-100 where as 6 and 8 mM CHAPSO concentration was required for this type of secondary structure. Further addition of either of the detergents result in the induction of a-helix in both the IgGs as evident by helix specific peaks in the amide I region of FTIR and circular dichroism spectra. Tryptophan and 8-anilino-1-naphthalene-sulphonic acid (ANS) fluorescence confirmed changes in protein conformation upon addition of detergents. Maximum ANS binding at 0.12 mM TX-100 in both while 6 and 8 mM CHAPSO in bovine and human IgG respectively, indicate a compact "molten-globule"-like conformation. An increase addition of these detergents results in the burial of hydrophobic patches of both IgG owing to aggregation. Presence of aggregates at 0.2 and 0.16 mM TX-100 and 8 and 9 mM CHAPSO, for bovine and human IgG respectively, was further confirmed by reduction in ANS fluorescence, dynamic light scattering study, thioflavin T fluorescence and congo red absorbance. (C) 2013 Elsevier B.V. All rights reserved.
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页码:151 / 160
页数:10
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