Delayed removal of N-terminal methionine from nascent globin chains in sickle-cell anemia reticulocytes.

Synthetic alphaT1 and betaT1, the N-terminal tryptic peptides of alpha-chain and beta-chain of hemoglobin, and MetalphaT1 and MetbetaT1, peptides in which N-terminal methionyl residues are peptide-bonded to alphaT1 and betaT1, were prepared by the solid-state method of Merrifield. These synthetic peptides were used to establish conditions for chromatographic purification and analysis. When tryptic digests of nascent globin chains from rabbit and sickle-cell anemia reticulocytes incubated with (35)S- and (3)H-labeled amino acids were analyzed, radioactivity not present in tryptic digests of labeled hemoglobin appeared at the elution positions of MetalphaT1 and MetbetaT1. The fraction of nascent chains with N-terminal methionine was higher in sickle-cell anemia reticulocytes than in rabbit reticulocytes. If rate of peptide-chain elongation in polysomes is uniform, nascent human chains must attain a greater length before removal of the initial methionyl residue. Length of nascent chain at time of removal was calculated from two independent sets of data, one obtained from [(35)S]methionine incorporation into MetalphaT1, MetbetaT1, alphaT5, and betaT5, and the other obtained from [(3)H]lysine and [(3)H]valine incorporation into MetbetaT1 and betaT1.