hrvatski

Tkivno specifični stanični i izvanstanični profil glikoproteina u staničnim linijama šećerne repe

Glikoproteini su velika grupa proteina koji imaju šećernu komponentu na N ili C kraju. Uključeni su u važne biološke procese kao što su razvoj, rast i preživljavanje organizma. Tkivne linije šećerne repe: N (normalna, ovisna o hormonima), HO (prilagođena, organogena) i HNO (prilagođena, neorganogena) već su morfološki i biokemijski opisane. Utvrđeno je da se nalaze na različitim stupnjevima razvitka. Međutim, detaljna glikoproteinska slika nije analizirana. U eksperiment smo dodatno uvrstili tri transformirane linije: T, Tr i Tw. U radu smo koristili lektine da bi karakterizirali šećernu komponentu staničnih i izvanstaničnih glikoproteina staničnih linija šećerne repe. Proteine razdvojene SDS-PAGE-om te prenesene na nitroceluloznu membranu, tretirali smo s lektinima SNA, MAA, RCA, UEA i ConA. Stanični glikoproteini su locirani između 49 i 55 kDa te između 22 i 33 kDa. Potonji nisu reagirali s ConA. Glikoproteini od 22 do 30 kDa detektirani pomoću UEA, bili su vidljivi samo u HO, Tr i Tw linijama. Sijalinska kiselina, glikanska komponenta koja dosad nije otkrivena u biljkama, detektirana je u svim staničnim linijama lektinima SNA i MAA. Vrpca od 60 kDa nije otkrivena U T liniji dok su vrpce od 24 – 29 kDa nedostajale u HNO i Tr liniji. Izvanstanični glikoproteini od 32 i 70 kDa bili su zajednički u svim linijama osim linije HNO. Neoplastična i dediferencirana linija HNO izlučivala je više proteina od drugih linija te su specifični glikoproteini otkriveni na 67, 42 i 40 kDa. Glikoproteini su povezani s fenotipom i s promjenama na razvojnoj razini. Glikoproteinska slika izvanstaničnih proteina pokazala je više razlika među tkivima nego slika staničnih. Determinacija specifičnih glikoproteina će se nastaviti. Glycoproteins are large group of proteins that have sugar component attached on the N or C terminal domain. They are involved in crucial biological events as development, growth, function and survival of an organism. Sugarbeet callus lines: N (normal, hormone dependent), HO (habituated organogenic) and HNO (habituated non - organogenic) were already morphologically and biochemicaly characterised, and found to be in different stages of differentiation. However, detailed glycoprotein patterns of these lines were not established. Three transformed lines: T, Tr and Tw were also included in the experiment. Lectins were used to characterise a sugar component of cellular and extracellular glycoproteins. Proteins separated by SDS-PAGE and immobilized on nitrocellulose membrane were probed with SNA, MAA, RCA, UEA and ConA lectins. For cellular glycoproteins, common bands were resolved in the range between 49-55 kDa and between 22-33 kDa. The latter bands did not react with ConA. Glycoproteins of 22-30 kDa detected with UEA were expressed in HO, Tr and Tw lines only. Sialic acid, a glycan component so far not reported in plants was detected in all tissue lines with SNA and MAA. The 60 kDa band was missing in T and 24 – 29 kDa bands were missing in HNO and Tr line. The extracellular 32 and 70 kDa glycoproteins were detected in all lines except in HNO. The HNO, neoplastic-dedifferentiated cell type excreted more proteins than other tissues and it had specific glycoproteins detected at 67, 42, 40 kDa. Glycoproteins are related with the phenotype and to changes in differentiation state. Extracellular glycoprotein patterns showed more differences between tissues than the cellular ones. Determination of specific glycoproteins will be the further step.

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