The reproduction of animals is a means to maintain their species. The golden hamsters are seasonal breeders, and their reproduction is regulated by the photoperiod. The shifts of the sexual functions imply the changes of the protein expression, and those are reflected by the proteome. Thus the present study was to examine via two dimensional polyacrylamide gel electrophoresis (2d PAGE), the physiological changes and the alterations of protein expressions in hypothalamus upon the reproductive situation related to the pineal gland. Among the pineal intact animals, the reproductive functions were sustained in animals housed in long photoperiod (LP) but regressed in animals housed in short photoperiod. And those pinealectomized animals showed high sexual activities regardless of photoperiod. Ultimately they were branched into dichotomy, sexually active and inactive animals. Apart from the changes of physiological parameters upon the reproductive conditions, there were obvious differences in proteins extracted from the hypothalamus. The hypothalamus of LP animals presented high levels of enzymes which are involved in the production of energy, glycolysis and Krebs cycle. The increased energy might be related to the GnRH synthesis in hypothalamus and indirectly to the constant cell divisions in spermatogenesis. Taken together, the impacts of the photoperiodic information controlling reproduction could be observed through 2d PAGE. Therefore, the present results suggest the potential of biomarkers collectively to diagnose the fertility and the infertility by way of proteomics in organs with regard to the reproductive system, further could be applied to diagnose various diseases.

and distribution of seed storage proteins are responsible for the quality of soybean and seed development. Among storage proteins, lipoxygenase isoforms (Mw. ~97 kDa) play a major role in the distinct bean flavor during storage. In this study, we compared three soybean elite cultivars viz., JIMPUM, JINPUM2 and TANMI2 (lipoxygenase null mutants, originated from Japan) along with WILLIAMS 82 (control plant, USA) to determine the seed storage proteins by proteomic approach. Phenotype of the mature seeds showed the variation in seed coat, color and appearance. Total seed proteins of the above cultivars were subjected to two dimensional gel electrophoresis (2-DE). The resulted protein profiling showed the intensity of the different quantitative spots varied among the four cultivars. We are now investigating by using other proteomic tool and the resulted difference in proteins may helpful in quality improvement or the functional roles in the seed development.

To better understanding the function of the luminal sub-organelles within the thylakoid network, we have carried out a systematical analysis and identification of the lumenal proteins in the thylakoid of wheat by using Tricine / 1D-PAGE, and LTQ-ESI-FTICR mass spectrometry followed by SWISS-PROT database searching. We isolation and fractionation these membrane from fully developed wheat leaves using a combination of differential and gradient centrifugation couple to high speed ultra-centrifuge. After collecting all proteins to eliminate possible same proteins, we estimated that there are 407 different proteins including chloroplast, chloroplast stroma, lumenal, and thylakoid membrane proteins excluding 20 proteins, which were identified in nucleus, cytoplasm and mitochondria. A combination of these three programs (PSORT, TargetP, and TMHMM) was found to provide a useful tool for evaluating chloroplast localization, transit peptide, transmembranes, and also could reveal possible alternative processing sites and dual targeting. Finally, we report also sub-cellular location specific protein interaction network using Cytoscape software, which provides further insight into the biochemical pathways of photosynthesis. The present work helps understanding photosynthesis process in wheat at the molecular level and provides a new overview of the biochemical machinery of the thylakoid in wheat.

Buckwheat is one of the traditional crops and has become a renewed target of interest or a popular crop as a healthy foodstuff, because it is a good source of cereal protein which is rich with essential amino acids. However, what is critical to our health is that buckwheat contains proteins which cause a allergy. Buckwheat allergy resulting from ingestion is caused by the storage proteins in the grain with molecular weights ranging from 15, 22, 35, 39 and 50 kDa proteins of the inner fractions to low, and there were clear differences in the protein compositions between the inner and outer buckwheat flour fractions. A major allergenic protein of buckwheat is Fag e 1 with molecular weight 22 kDa (BW22KD). Buckwheat allergy is an immunoglobulin E (IgE)-mediated hypersensitive response capable of causing anaphylactic shock. Buckwheat seeds were dissected to endosperm and embryo. From each fraction we extracted proteins and analyzed extracts by SDS-PAGE and 2-DE. On electrophoregrams of endosperm proteins, 6 intense bands were detected. The most intense corresponded to molecular weights ranging from 54 to 65 kDa. These proteins have been reported not to be allergenic. We show here that the allergenic buckwheat seed proteins are found only among embryo proteins. No allergenic proteins were found in the buckwheat endosperm. The results presented here lead to the proposal that patients with hypersensitivity to buckwheat flour should use only fine flour from buckwheat endosperm, as this fraction contains no allergenic proteins. At present, specific protein spots will be selected and in-gel digested for MALDI-TOF-TOF/MS analysis.

The C3HC4 zinc RING finger proteins seem to be a family of protein-protein interactions. Little is information regarding the role of the C3HC4 zinc RING finger proteins in rice plant. We have attempted to assess their genome localization, phylogenetic relationship and expression patterns of members via in silico analysis as well as semi-quantitative RT-PCR. A total of 132 genes encoding C3HC4 zinc RING finger proteins appear to be distributed over 12 rice chromosomes, reflecting evolutionary dynamics of the rice genome, e.g. whole genome duplication and tandem duplications. A genome-wide dataset including 155 gene expression omnibus sample (GSM) plates evidenced a high degree of functional specialization of the rice C3HC4 zinc RING finger proteins, especially during developmental stages and against abiotic stresses. We have retrieved co-expression genes with each of the rice C3HC4 zinc RING finger proteins, probably providing some clues on specialized functions of individual genes. Expression patterns of 13 co-expression genes with one gene encoding C3HC4 zinc RING finger protein (Os04g51400) against salt and dehydration stresses were evaluated in crown tissues and leaf tissues, evidencing highly similar patterns among members. These findings might provide clues to shed further light on comprehensive functions of C3HC4 zinc RING finger proteins.

An attempt was made to link rice embryo proteins to DNA sequences and to understand their functions. One hundred of the 700 spots detected on the embryo 2-DE gels were microsequenced. Of these, 28% of the embryo proteins were matched to DNA sequences with known functions, but 72% of the proteins were unknown in functions as previously reported (Woo et al. 2002). In addition, twenty-four protein spots with 100% of homology and nine with over 80% were matched to ESTs (expressed sequence tags) after expanding the amino acid sequences of the protein spots by Database searches using the available rice EST databases at the NCBI (http://www/ncbi.nlm.nih.gov/) and DDBJ (http://www.ddbj.nig.ac.jp/). The chromosomal location of some proteins were also obtained from the rice genetic map provided by Japanese Rice Genome Research Program (http://rgp.dna.affrc.go.jp). The DNA sequence databases including EST have been reported for rice (Oryza sativa L.) now provides whole or partial gene sequence, and recent advances in protein characterization allow the linking proteins to DNA sequences in the functional analysis. This work shows that proteome analysis could be a useful tool strategy to link sequence information and to functional genomics.