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Anomalous electrophoretic behavior of a very acidic protein: Ribonuclease U2
Author(s) -
GarcíaOrtega Lucía,
De los Ríos Vivian,
MartínezRuiz Antonio,
Oñaderra Mercedes,
Lacadena Javier,
Martínez del Pozo Álvaro,
Gavilanes José G.
Publication year - 2005
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200500261
Subject(s) - electrophoresis , ribonuclease , molecular mass , chemistry , dimer , gel electrophoresis , chromatography , biochemistry , enzyme , rna , organic chemistry , gene
Ribonuclease U2 is a low‐molecular‐weight acidic protein with three disulfide bridges. This protein displays an anomalous electrophoretic behavior on standard SDS‐PAGE. The electrophoretic mobility of the nonreduced protein roughly corresponds to its molecular mass while the migration of the reduced protein would be in accordance with the expected molecular mass of the protein dimer. This study reveals that the protein does not bind SDS under the SDS‐PAGE conditions, its electrophoretic mobility being only determined by its electrostatic charge and hydrodynamic properties. In addition, the nonreduced protein cannot be blotted to a membrane. Unfolding of the protein upon reduction of its disulfide bridges enables electrotransference to membranes due to a restricted diffusion along the electrophoresis gel.