Determination of pore/protein size via electrophoresis and slit sieve model.
We used electrophoresis for three purposes: (i) estimation of the mean pore size of polyacrylamide gels via measuring electrophoretic mobility of globular proteins of known sizes in combination with simple sieve (cylindrical and slit) models; (ii) determination of the average size of protein molecules (native or denatured) by the use of the same models; (iii) monitoring the changes in molecular dimensions of proteins in the course of their denaturation. Both models yield results that are in good agreement with those found via the more elaborate techniques (considering the principal differences involved). The approach provides a direct and convenient way of monitoring the variations in protein sizes during the course of their denaturation in gels having a gradient of denaturants, and possibly the number of conformational states involved in the process, a facet that is quite unique and useful. The simpler slit model seems to yield better results in the latter case and is moreover supported by the recently reported data on electrophoresis of DNA molecules through the 1 microm slits of a microbrush matrix made of micropillars arranged in a hexagonal lattice.
Sarbolouki
Mohammad N
MN
Institute of Biochemistry and Biophysics, Tehran University, Tehran, Iran. sarbol@ibb.ut.ac.ir
Mahnam
Karim
K
Rafiee-Pour
Hossain-Ali
HA
D004586:Electrophoresis / Q000379:methods; D007202:Indicators and Reagents; D008970:Molecular Weight; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification; D015203:Reproducibility of Results; D014508:Urea
D007202:Indicators and Reagents; D011506:Proteins; D014508:Urea