N-acetylneuraminic acid, phosphate and thiol groups of pyruvate kinase isoenzymes from Morris hepatoma 7777 and normal rat liver.
The highest amount of N-acetylneuraminic acid (AcNeu) was found in pyruvate kinase isoenzyme L from normal rat liver (24 moles/mole of enzyme tetramer), with the highest electrophoretic mobility. On the other hand, isoenzyme M2 from Morris hepatoma 7777, with the lowest electrophoretic mobility, had the lowest AcNeu content (5 moles/mole of enzyme tetramer). This tumour isoenzyme M2 of pyruvate kinase was, however, characterised by the highest phosphate content (12 moles/mole protein), in comparison to isoenzyme L (3 moles/mole protein) or normal liver isoenzyme M2 (6 moles/mole protein). This could indicate a regulatory change caused by reversible enzyme phosphorylation and dephosphorylation or sialization and desialization. Despite these differences, the sum of the two negatively charged residues was lower in tumour pyruvate kinase isoenzyme M2, with the slowest migration rate, than in normal rat liver isoenzyme M2. Moreover, isoenzyme M2 from tumour material, in comparison with isoenzyme M2 from normal rat liver, had a twice as high content of thiol groups (20 moles/mole protein), especially of free and superficially located ones, than the isoenzyme M2 from normal liver (10 moles/mole protein). This may explain abnormal susceptibility of tumour isoenzyme M2 to stereospecific inhibition by exogenous L-cysteine, and indicate genetically dependent changes in amino-acid content of tumour enzyme which take place during cell tumourigenic transformation.
Ignacak
J
J
Department of General Biochemistry, Collegium Medicum, Jagiellonian University, Cracow, Poland.
Gumińska
M
M
D000818:Animals; D003545:Cysteine / Q000032:analysis; D003600:Cytosol / Q000201:enzymology; D004220:Disulfides / Q000032:analysis; D004586:Electrophoresis; D007527:Isoenzymes / Q000032:analysis*; D008099:Liver / Q000201:enzymology* / Q000378:metabolism; D008114:Liver Neoplasms, Experimental / Q000201:enzymology* / Q000378:metabolism; D009368:Neoplasm Transplantation; D010710:Phosphates / Q000032:analysis*; D011770:Pyruvate Kinase / Q000032:analysis*; D051381:Rats; D011915:Rats, Inbred BUF; D012680:Sensitivity and Specificity; D012794:Sialic Acids / Q000032:analysis*; D013438:Sulfhydryl Compounds / Q000032:analysis*
D004220:Disulfides; D007527:Isoenzymes; D010710:Phosphates; D012794:Sialic Acids; D013438:Sulfhydryl Compounds; D011770:Pyruvate Kinase; D003545:Cysteine