Increased membrane-protein methylation in hereditary spherocytosis. A marker of cytoskeletal disarray.
Protein carboxyl methyltransferase of type II selectively recognizes L-isoaspartyl and D-aspartyl residues spontaneously occurring in proteins and peptide substrates. Membrane protein methylation levels increase with erythrocyte aging in circulation, in parallel with the spontaneous formation of abnormal aspartyl sites, due to protein intrinsic instability. We found that enzymic methyl esterification of erythrocyte membrane proteins in hereditary spherocytosis, a model of cytoskeletal disarray, is significantly increased compared to normal red blood cells. This cannot be explained by an increase in mean age of spherocytes, which are on the contrary significantly younger than control cells. No differences in cytosolic methyltransferase specific activity, as well as in the intracellular concentrations of the methyl donor adenosylmethionine and/or of the methylation inhibitor adenosylhomocysteine were observed. We identified bands 2.1, 4.1 and 4.2 as the main targets for increased methylation, whose levels were correlated with the degree of spectrin deficiency associated with this anemia. Our findings indicate that membrane-protein methyl esterification represents a marker of membrane structural alteration in vivo in spherocytosis. We hypothesize that either an increased accessibility of methylation sites normally not available to the methyltransferase, or accelerated formation of methyl-accepting sites in membrane proteins are present in spherocytosis.
Ingrosso
D
D
Institute of Biochemistry of Macromolecules, Second University of Naples, Italy.
D'Angelo
S
S
Perna
A F
AF
Iolascon
A
A
Miraglia del Giudice
E
E
Perrotta
S
S
Zappia
V
V
Galletti
P
P
D003599:Cytoskeleton / Q000378:metabolism*; D004910:Erythrocyte Membrane / Q000378:metabolism*; D004951:Esterification; D006801:Humans; D008565:Membrane Proteins / Q000378:metabolism*; D008745:Methylation; D011496:Protein Methyltransferases / Q000378:metabolism; D012435:S-Adenosylhomocysteine / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000378:metabolism; D013049:Spectrin / Q000172:deficiency; D013103:Spherocytosis, Hereditary / Q000378:metabolism*
D008565:Membrane Proteins; D013049:Spectrin; D012436:S-Adenosylmethionine; D012435:S-Adenosylhomocysteine; D011496:Protein Methyltransferases