[en] We introduce a simple theoretical approach for an equilibrium study of proteins with known native-state structures. We test our approach with results on well-studied globular proteins, chymotrypsin inhibitor (2ci2), barnase, and the alpha spectrin SH3 domain, and present evidence for a hierarchical onset of order on lowering the temperature with significant organization at the local level even at high temperatures. A further application to the folding process of HIV-1 protease shows that the model can be reliably used to identify key folding sites that are responsible for the development of drug resistance

[en] Trypsin and chymotrypsin inhibitor activities of safflower oilcake were studied before and after irradiation. The various doses to which samples were exposed ranged from 7 Gy to 10 kGy. The trypsin inhibitor is inactivated at 42 Gy, whereas the chymotrypsin inhibitor remains active, even at the much higher dose of 10 kGy. Thein vitro digestibility values also showed a significant improvement after irradiation. Exposure to a low dose of 42 Gy is sufficient to improve the nutritional value of the oilcake

[en] Full text: α-Trypsin is a serine-protease with a polypeptide chain of 223 amino acid residues and six disulfide bridges. It is a globular protein with predominance of antiparallel β-sheet secondary structure and it has two domains with similar structures. In the present work, a stability study of α-trypsin in the acid pH range was performed and physical-chemical denaturation parameters were measured by using differential scanning calorimetry (DSC). The α-trypsin has a shelf-life (t_95%) of about ten months at pH 3.0 and 4 deg C and its hydrolysis into the Ψ-trypsin isoform is negligible during six months as monitored by mass spectrometry (Micromass Q-ToF). The observed ΔH_cal/ΔH_vH ratio is close to unity for α-trypsin denaturation, which suggests the occurrence of a two-state transition, devoid of molten-globule intermediates. At pH 3.0, α-trypsin unfolded with T_m 325.9 K and ΔH= 99.10 kcal mol^-1, and the change in heat capacity between the native and unfolded forms of the protein was estimated to be 1.96 ± 0.18 kcal mol^-1 K^-1. The stability of α-trypsin calculated at 298 K and at pH 3.0 was ΔG_U = 6.10 kcal mol^-1. These values are in the range expected for a small globular protein. These results show that the thermodynamic parameters for unfolding of β-trypsin do not change substantially after its conversion to α-trypsin

[en] Highlights: • The elastic modulus of the enzymatically digested LDL was evaluated using AFM. • The LDL elastic modulus was decreased not by proteases but by phospholipase A₂. • The LDL size was unaffected by proteases or PLA₂. Oxidation of low-density lipoproteins (LDLs) induces development of cardiovascular disease. Recently, reports of studies using atomic force microscopy (AFM) have described that the elastic modulus of metal-induced oxidized LDLs is lower than the modulus before oxidation. However, the mechanisms of change of the elastic modulus have not been well investigated. We postulated that disorder of the LDL structure might decrease the elastic modulus. This study measured the elastic modulus of LDLs before and after enzyme treatment with V8 protease, α-chymotrypsin, and phospholipase A₂. After LDLs were obtained from serum by ultracentrifugation, LDLs or enzyme-treated LDLs were physically absorbed. They were crowded on a mica surface. Although V8 protease and α-chymotrypsin did not induce the elastic modulus change, treatment with PLA₂ decreased the elastic modulus. The LDL particle size did not change during the enzyme treatment. Results suggest that disordering of the lipid structure of the LDL might contribute to the elastic modulus change. Results show that AFM might be a useful tool to evaluate disorders of complex nanoscale particle structures from lipids and proteins such as lipoproteins.

No abstract available

No abstract available

[en] The effects of heat treatment and enzymic digestion on the antigenic reactivity of ovomucoid were studied. This reactivity was mainly examined by a solid phase competitive radioimmunoassay. The trypsin inhibitory activity was also measured to elucidate its relationship to the antigenic reactivity. The antigenic reactivity and trypsin inhibitory activity diminished in parallel with increase of heating time. However, enzymic digestion caused a more rapid decrease in antigenic reactivity than in trypsin inhibitory activity. From these results, it is suggested that the structure of the antigenic determinants is destroyed in a similar manner to that of the trypsin inhibitory active site when ovomucoid is heated, while the former structure is destroyed more easily than the latter by enzymic digestion

No abstract available

[en] The newly developed magnetic micro- and nanoparticles with defined hydrophobicity and porosity were used for the preparation of magnetic enzyme reactors. Magnetic particles with immobilized proteolytic enzymes trypsin, chymotrypsin and papain and with enzyme neuraminidase were used to study the structure of heterogeneous glycoproteins. Factors such as the type of carrier, immobilization procedure, operational and storage stability, and experimental conditions were optimized

No abstract available