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Phytohemagglutinin purification: A general method involving affinity and gel chromatography

1973; Wiley; Volume: 34; Issue: 1 Linguagem: Inglês

10.1016/0014-5793(73)80702-1

1873-3468

Wolfgang G. Bessler, Ido Goldstein,

Plant Stress Responses and Tolerance

Pr0~e~nz agglutinating erythr~cytes and other types of cells have been studied since 1888 when Stil]mark ,ob~er~,ed that extracts fr,em Ric~n~s eommzmis clumpe,d ,led bleed ,cells [1].H~.w,evez. it is only zince the experiments of Boyd I2. 3] in 3945 on the zpecificity of" a large number of phytohemagglutinins 0eetins) I4] tha't these seed p;oteins became a subject ef g,en,erM ire'erect ~5].Research on teedns has ,increased marked-]y within the hs* ~ew yea,~s after it was shown thai LtlCy 5n~ezac/with no~i and cancerous cells 16--9].Beemuse of tile remarkable interest gen,era~,ed in ~he many ~xrsiqn,e biological ,properties of leeti,ns and &e ,large number of p~oeed~ a.s requi~e,d to purify 5n,dividuz.lphytohemagglutinins0 we have tried to develop a rn,oxe gen,eral me/hod for fla~ir pnri~a~ion (of.~[lO])-For this purpose, we have ~ak,en a,dvan'talge of ~,e dem,onslra~e,d ,eapaeity of ooneanavalin A to i'ntrerael, ~h most phytoh,emagg~lu'tinins of the family Leg~min,osae I] 1] ,and of th~ tfigh r,e~olving powe~ of long crolunms of Sephadex G-200 ~aperfine gel.This communieation describes ,.the purificNion of foul lee-~ins ,emp],oying ,essentially three pu:ri:ficat~on steps: 5) '(NH~)2 SO~ fza,etiona~c-n ,of the crude .extracts,Yx) affmd~y chlomat,ogra~ on Con A-Bepharose and iii) .¢hroma't,ography,on Eephadex 'G-200 supelfme get.The plant a ggludni'ns in~esligated were from lima bean (Phaseolus luna.s),soy ~ean ,(Glycine max), wax bean ,(Phas eo lu~ ~,ulg aris) and ~ared eira ea sfm p li cif o li~ .~re~ liminary results on attempted purLqealion ,Of ,O'l~leI leetins a~e ~ls0 reported. 58