Limited tryptic hydrolysis of pea legumin: molecular mass and conformational stability of legumin-T.
The investigation of hydrodynamic and thermodynamic properties and the determination of the molecular mass of legumin-T, the product of limited tryptic hydrolysis of the 11-S-globulin from pea seeds, was carried out to ascertain the structural relationship to globulin-T's from other legumin-like proteins. The obtained legumin-T preparation has a molecular mass M(W)=260+/-10 kDa and M(S,D)=270+/-20 kDa. The secondary structure of legumin-T is characterised by a high percentage of beta-sheet conformation, comparable to that of native legumin and a reduced percentage of helical conformation. The conformational stability of legumin-T evaluated by equilibrium unfolding in the presence of guanidinium chloride was only slightly reduced in comparison to the native legumin, whereas the calorimetrically determined denaturation enthalpy and Gibbs energy of denaturation were found to be increased for legumin-T. These physicochemical properties are very similar to those of faba bean legumin-T.
Schwenke
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KD
Institut für Angewandte Proteinchemie e.V., c/o Biologische Bundesanstalt, Stahnsdorfer Damm 81, D-14532, Kleinmachnow, Germany. food.proteins@prochem.pm.shuttle.de
Henning
T
T
Dudek
S
S
Dautzenberg
H
H
Danilenko
A N
AN
Kozhevnikov
G O
GO
Braudo
E E
EE
D002151:Calorimetry; D002847:Chromatography, Agarose; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D005916:Globulins / Q000737:chemistry; D019791:Guanidine / Q000737:chemistry; D006868:Hydrolysis; D008027:Light; D018532:Pisum sativum; D010940:Plant Proteins / Q000737:chemistry*; D011487:Protein Conformation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature; D013816:Thermodynamics; D014357:Trypsin / Q000737:chemistry* / Q000378:metabolism*; D014867:Water / Q000737:chemistry; D000097793:Legumins
D005916:Globulins; D019791:Guanidine; D010940:Plant Proteins; D014357:Trypsin; D014867:Water; D000097793:Legumins