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Labeling of pigeon erythrocyte membrane proteins by low level reductive methylation

Labeling of pigeon erythrocyte membrane proteins by low level reductive methylation

Analytical Biochemistry 117(2): 459-465

The reductive methylation procedure was adapted for 3H-labeling of membrane proteins using pigeon erythrocyte membrane. Usably high 3H incorporation into protein was obtained, e.g., 28 .mu.Ci/mg protein with 83 nmol (input) H2CO/mg protein, B3H4- at 10 Ci/mmol, and a B3H4-/H2CO ratio of 0.34. With this low H2CO/protein ratio, methylation did not perturb ATP-dependent 45Ca2+ uptake, Na+-dependent [14C]glycine uptake, membrane vesicle sealing, or isoelectric focusing patterns of methylated membrane proteins. The labeled membrane proteins were shown to be good tracers for the unlabeled proteins by using 2-dimensional isoelectric focusing .times. sodium dodecyl sulfate gel electrophoresis.

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Accession: 005787035

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PMID: 7325375

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