Preparation of ribsomal subunits by gel filtration

Bhoolia, Deena
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An attempt was made to separate ribosomal subunits by gel filtl'ation on Trisacryl GF2000 and Sepharose 4B. Trisacryl GP~2000,a sYllthet'ic gol, , separated rat .ljver rlbosonal subunits orl 'f~'135,em column with a resolution of ""Or3, resulting 'in <'1" 60 rJ impm''ity 'of each of the \~ subunits. subunits" were not resolved. Sepharose 4131 an agarose based gol, separated " the subunits by adsorpttcn chromatography rathr.r than 'i>.V () Q ':) At 4°C, the 405 .subunf ts were eluted with a kd () ruO,:~9( but the GOrf' $ubud"its adsorbed to th~.'g,~Jl and were eluted I; /) when the temperature of "the column was increas~d to 250C..3SoC. (' ,. ('-. This edsorpt ion phenonenon seems to b{~ tl propert~ of a 11 agllrose -: \') ~\:; based gels studied here, includ'ing Sepharose 2.B and .seph'ato~eoBt arid is exc 1us iva to !,mamm'i'lan r ibosOIntrt subuni ts • Anal,ys1S 0'(" the , u subunits by in vitro r14C]polypheny1alanine sy'nthes'is showed OCI " ,,'c. /\ I 7', " diff'erence 'in the act lvtt ies of dbosoma'J ~ubunH~ p~epdrf#d by ;/'/ ' grndient centrifugation or by 5epharos,e cni"OmatogNPPY;' Analy~ii ~~of " (~ (( the subunits by ~crylamidec'ugarose coU)po5it~) gs1s resulted ,in the '/ resolution ,.:n"f subunits isolated fr'oill lower organisms in o II non-denaturlnq !Jcl systems and SI,lbut1its from m«mm~nan'tissue in II /'\';1 Ga'! f'iltrat'ion does tyffer a 5t!i:l;~'ble metrKld'¥or the pr~p;n~¥tiol1of <::) . I \) \\ I) ribosoma 1 'subunits I but only' if 'the act~orption JH'OrJ&~"t'Jo,s of ,) (,) ':' o ,1) ,{
A Oiss{!y'tation $uhm'itted in fulfilment of the r~;~airements, for' the degree of f·1aster· oi~'j Science at the University of the ~titwatersrand ' Juhanoasbul"g January 1991