Resistance of soybean trypsin inhibitor (Kunitz) to denaturation by guanidinium chloride.

Leach, Bonnie Strayer; Fish, Wayne W.

doi:10.1016/s0021-9258(19)63337-5

Cited by 22 publications

(3 citation statements)

References 28 publications

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“…a Stokes radius of 6.4 ±0.1 nm (this corresponds to an [tj] of ~29.5 mL/g). This value for the Stokes radius is in good agreement with the value of 6.2 nm that can be estimated for a randomly coiled polypeptide having the chain length and disulfide bond pattern of antithrombin (Tanford, 1968;Leach & Fish, 1977; Peterson et al, 1979). In keeping with the latter result, sedimentation equilibrium measurements of disulfidecross-linked BAT in 6 M GdmCl yielded a molecular weight of 53 500 which indicates that the denatured protein is monomeric under these conditions.…”

Section: Resultssupporting

confidence: 86%

Denaturation behavior of antithrombin in guanidinium chloride. Irreversibility of unfolding caused by aggregation

Fish¹,

Danielsson²,

Nordling³

et al. 1985

Biochemistry

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The structural stability of the protease inhibitor antithrombin from bovine plasma was examined as a function of the concentration of guanidinium chloride (GdmCl). A biphasic unfolding curve at pH 7.4, with midpoints for the two phases at 0.8 and 2.8 M GdmCl, was measured by far-ultraviolet circular dichroism. Spectroscopic and hydrodynamic analyses suggest that the intermediate state which exists at 1.5 M GdmCl involves a partial unfolding of the antithrombin molecule that exposes regions of the polypeptide chain through which slow, intermolecular association subsequently takes place. The partially unfolded molecule can be reversed to its fully functional state only before the aggregation occurs. Upon return of the aggregated state to dilute buffer, the partially unfolded antithrombin remains aggregated and does not regain the spectroscopic properties, thrombin-inhibitory activity, or heparin affinity of the native inhibitor. This behavior indicates that the loss of the functional properties of the proteins is caused by the macromolecular association. Comparative experiments gave similar results for the human inhibitor. Analyses of bovine antithrombin in 6 M GdmCl indicated that the second transition reflects the total unfolding of the protein to a disulfide-cross-linked random coil. This transition is spectroscopically reversible; however, on further reversal to dilute buffer, the molecules apparently are trapped in the partially unfolded, aggregated, intermediate state. The results are consistent with the existence of two separate domains in antithrombin which unfold at different concentrations of GdmCl but do not support the contention that the thrombin-binding and heparin-binding regions of the protein are located in different domains [Villanueva, G. B., & Allen, N. (1983) J. Biol. Chem. 258, 14048-14053].

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Section: Resultssupporting

confidence: 86%

Denaturation behavior of antithrombin in guanidinium chloride. Irreversibility of unfolding caused by aggregation

Fish¹,

Danielsson²,

Nordling³

et al. 1985

Biochemistry

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“…The two disulfides bridges between Cys 39 and Cys 86 and between Cys 138 and Cys 145 are solvent-exposed and are essential for inhibitory function. The protein has been classified in β-II or the disordered class of all antiparallel β-sheet proteins on the basis of characteristic circular dichroism (CD) spectra. , The most interesting features of the molecule reported are its reversibility of thermal denaturation, resistance to chemical denaturants, and stability toward acidic conditions similar to that of stomach. − …”

Section: Introductionmentioning

confidence: 99%

Influence of γ-Radiation on the Structure and Function of Soybean Trypsin Inhibitor

Mallikarjunan

Marathe

Deshpande

et al. 2012

J. Agric. Food Chem.

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Soybean trypsin inhibitor (STI) is a known antinutrient and food allergen present in soybean. γ-Radiation has the potential to inactivate the TI protein. However, a systematic study on the influence of different moisture levels during γ radiation on structure and function of the molecule has not been reported. Pure STI was irradiated up to 200 kGy, in dry state, with 50% moisture and in aqueous solution. The radiation damage in molecular structure was assessed using, SDS-PAGE, size exclusion chromatography, fluorescence measurement, and circular dichroism, while functional damage was assessed by the TI assay. In aqueous solution, both the structure and function of TI were almost destroyed at the 10 kGy dose. While with 50% moisture and in dry state, the loss in functional and structural attributes was discernible only at 30 and 100 kGy, respectively. The TI activity was found to be unaffected in dry and soaked seeds of soybean as well as other legumes up to irradiation doses of 100 and 50 kGy, respectively.

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“…Esta é uma segunda característica marcante desta classe de proteínas, a resistência a agentes caotrópicos, como já descrita para diversos inibidores, dentre eles o inibidor de (GRUEN ., 1984), o qual apresentou uma elevada similaridade com EATI, baseado na composição global de aminoácidos. Estudos de desnaturação de STI por cloreto de guanidina conduzidos Leach & Fish (1977) concluíram que o desenovelamento completo deste inibidor acontece após 14 dias de incubação com 6…”

Section: S-i S-iiunclassified

Aspectos funcionais e estruturais do inibidor de tripsina de Entada acaciifolia

Oliveira¹,

Macedo²

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Resistance of soybean trypsin inhibitor (Kunitz) to denaturation by guanidinium chloride.

Cited by 22 publications

References 28 publications

Denaturation behavior of antithrombin in guanidinium chloride. Irreversibility of unfolding caused by aggregation

Denaturation behavior of antithrombin in guanidinium chloride. Irreversibility of unfolding caused by aggregation

Influence of γ-Radiation on the Structure and Function of Soybean Trypsin Inhibitor

Aspectos funcionais e estruturais do inibidor de tripsina de Entada acaciifolia

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