Proteomic analysis of the photosystem I light-harvesting antenna in tomato (Lycopersicon esculentum).
Until now, more genes of the light-harvesting antenna of higher-plant photosystem I (PSI) than proteins have been described. To improve our understanding of the composition of light-harvesting complex I (LHCI) of tomato (Lycopersicon esculentum), we combined one- and two-dimensional (1-D and 2-D, respectively) gel electrophoresis with immunoblotting and tandem mass spectrometry (MS/MS). Separation of PSI with high-resolution 1-D gels allowed separation of five bands attributed to proteins of LHCI. Immunoblotting with monospecific antibodies and MS/MS analysis enabled the correct assignment of the four prominent bands to light-harvesting proteins Lhca1-4. The fifth band was recognized by only the Lhca1 antibody. Immunodetection as well as mass spectrometric analysis revealed that these protein bands contain not only the eponymous protein but also other Lhca proteins, indicating a heterogeneous protein composition of Lhca bands. Additionally, highly sensitive MS/MS allowed detection of a second Lhca4 isoform and of Lhca5. These proteins had not been described before on the protein level in higher plants. Two-dimensional gel electrophoresis revealed an even more diverse composition of individual Lhca proteins than was apparent from 1-D gels. For each of the four prominent Lhca proteins, four to five isoforms with different isoelectric points could be identified. In the case of Lhca1, Lhca4, and Lhca3, additional isoforms with slightly differing molecular masses were identified. Thus, we were able to detect four to ten isoforms of each individual Lhca protein in PSI. Reasons for the origin of Lhca heterogeneity are discussed. The observed variety of Lhca proteins and their isoforms is of particular interest in the context of the recently published crystal structure of photosystem I from pea, which showed the presence of only four Lhca proteins per photosystem I. These findings indicate that several populations of photosystem I that differ in their Lhca composition may exist.
Storf
Stefanie
S
Institut für Allgemeine Botanik, Johannes Gutenberg-Universität Mainz, Müllerweg 6, 55099 Mainz, Germany.
Stauber
Einar J
EJ
Hippler
Michael
M
Schmid
Volkmar H R
VH
D000595:Amino Acid Sequence; D004586:Electrophoresis; D015151:Immunoblotting; D045342:Light-Harvesting Protein Complexes / Q000737:chemistry*; D018551:Solanum lycopersicum / Q000737:chemistry*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D045331:Photosystem I Protein Complex / Q000737:chemistry* / Q000302:isolation & purification; D020033:Protein Isoforms / Q000302:isolation & purification; D040901:Proteomics / Q000379:methods*; D020539:Sequence Analysis, Protein
D045342:Light-Harvesting Protein Complexes; D045331:Photosystem I Protein Complex; D020033:Protein Isoforms