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Napin from Brassica juncea: Thermodynamic and structural analysis of stability

Jyothi, T. C. and Sharmistha, Sinha and Sridevi, A. S. (2007) Napin from Brassica juncea: Thermodynamic and structural analysis of stability. Biochimica et Biophysica Acta, 1774. pp. 907-919.

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Abstract

The napin from Brassica juncea, oriental mustard, is highly thermostable, proteolysis resistant and allergenic in nature. It consists of two subunits – one small (29 amino acid residues) and one large (86 amino acids residues) – held together by disulfide bonds. The thermal unfolding of napin has been followed by differential scanning calorimetry (DSC) and circular dichroism (CD) measurements. The thermal unfolding is characterized by a three state transition with TM1 and TM2 at 323.5 K and 335.8 K, respectively; ΔCP1 and ΔCP2 are 2.05 kcal mol−1 K−1 and 1.40 kcal mol−1 K−1, respectively. In the temperature range 310–318 K, the molecule undergoes dimerisation. Isothermal equilibrium unfolding by guanidinium hydrochloride also follows a three state transition, N⇆I⇆U with ΔG1H2O and ΔG2H2O values of 5.2 kcal mol−1 and 5.1 kcal mol−1 at 300 K, respectively. Excess heat capacity values obtained, are similar to those obtained from DSC measurements. There is an increase in hydrodynamic radius from 20 Å to 35.0 Å due to unfolding by guanidinium hydrochloride. In silico alignment of sequences of napin has revealed that the internal repeats (40%) spanning residues 31 to 60 and 73 to 109 are conserved in all Brassica species. The internal repeats may contribute to the greater stability of napin. A thorough understanding of the structure and stability of these proteins is essential before they can be exploited for genetic improvements for nutrition.

Item Type: Article
Uncontrolled Keywords: Napin; Thermal stability; Brassica juncea; Association; Protein unfolding; Internal repeats
Subjects: 600 Technology > 08 Food technology > 19 Lipids-oils/fats > 04 Mustard seed
Divisions: Protein Chemistry and Technology
Depositing User: Food Sci. & Technol. Information Services
Date Deposited: 20 Nov 2008 09:47
Last Modified: 11 Oct 2018 08:16
URI: http://ir.cftri.res.in/id/eprint/8854

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