Identity of a Group of 13.5-20 kDa Polypeptides Whose Apparent Abundance Decreases during Plastid Development

The identity of a group of 13.5–20 kDa polypeptides whoseabundance decreases during the greening of intermittent-lightgrown barley seedlings was investigated. During greening therelative abundance of the 13.5–20 kDa polypeptides wasinversely related to that of LHC II, which had led others tosuggest a role of these polypeptides in the assembly of theLHC II and/or chloroplast development. The smallest 13.5 kDapolypeptide was identified as histone H4 by N-terminal sequencingof an internal peptide fragment produced by CNBr cleavage. Theentire group of 13.5–20 kDa polypeptides was thereafterverified to be nuclear histones by their similar mobility onSDS-PAGE to that of barley histones and immunoreactivity toyeast histone anti bodies. Their presence results from contaminationof plastid preparations by nucleosomes. Our results unequivocallysubstantiate earlier suggestions of others that polypeptidesoften found to contaminate immature plastids were of nuclearorigin. Methods to reduce or remove the histone contaminationwithout reduction in yield of thylakoids were developed so thattrue changes in the polypeptide content of thylakoids can bestudied during plastid development. (Received December 19, 1991; Accepted August 15, 1992)