Proteomic analysis of photosystem I components from different plant species

In this study, the photosystem I (PSI) highly hydrophobic proteins present within stroma lamellae of the thylakoid membrane were separated by RP-HPLC and identified either by in-solution trypsin digestion peptide fragment fingerprinting or by the close correspondence between the intact mass measurements (IMMs) and those expected from the DNA sequence. Protein identification performed by MS/MS was as reliable as IMMs. Thus, IMM is an easy and valid method for identifying proteins that have no PTMs. This paper reports the M(r) for all PSI proteins in ten different species, including those whose genes have not yet been cloned. Lhca5 was revealed unequivocally in four species, corroborating that it is indeed a protein belonging to the light-harvesting antenna of PSI. In all species examined, the product of the Lhca6 gene has never been revealed. Concerning core proteins, Psa-O has been revealed in three species; isoforms of Psa-D and Psa-E have been found in both monocots and dicots. Small proteins like Psa-I and Psa-J are well separated and identified. RP-HPLC produces reliable fingerprints and reveals that the relative amounts of PSI proteins appear to be markedly different.