SAMADI A K, MALEKNIA N. SEPARATION OF NONHISTONE HIGH MOBILITY GROUP (HMG) FROM HUMAN LYMPHOCYTES BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY. Med J Islam Repub Iran 1990; 4 (1) :65-70
URL:
http://mjiri.iums.ac.ir/article-1-1573-en.html
From the' Dept. of Biochemistry. Kashan University of Medical Sciences, Kashan
Abstract: (3883 Views)
The high mobility group (HMG) of nonhistone proteins have been
investigated using two high performance liquid chromatographic techniques
(HPLC). Reversed-phase HPLC under conditions of 50 mM triethylamine
adjusted to pH 2.2 with phosphoric acid (solvent A) and 95% acetonitrile in
water (solvent B) was used to separate proteins primarily on the basis of
differences in the overall hydrophobicity. Size exclusion HPLC under
conditions of two different solvents (A, 0.1 % trifluoroacetic acid 1FA B,
1.0% sodium dodecyl sulphate, SDS) was used to separate proteins. HMG
proteins from human lymphocytes were separated into the HMG 1, HMG 2,
HMG 14 and HMG 17 components. RP-HPLC is a proper method to resolve
all the human lymphocyte HMG-proteins. Size exclusion HPLC was employed
to resolve the HMG-protein subunits and determine their molecular
weights. Ideal SE-HPLC is not capable of resolving HMG 1 from HMG 2 or
HMG 14 from HM G 17 due to their molecular weight similarities. The purity
of protein fractions were examined by acetic acid-urea-triton X-100 gel
electrophoresis.