N␣ -Acetylation, catalyzed co-translationally with N ␣ -acetyltransferase (NAT), is the most common modifications of eukaryotic proteins. In yeast, there are at least three NATs: NAT1, MAK3, and NAT3. The 20 S proteasome subunits were purified from the normal strain and each of the deletion mutants, nat1, mak3, and nat3. The electrophoretic mobility of these subunits was compared by two-dimensional gel electrophoresis. Shifts toward the alkaline side of the gel and unblocking of the N terminus of certain of the subunits in one or another of the mutants indicated that the ␣1, ␣2, ␣3, ␣4, ␣7, and ␤3 subunits were acetylated with NAT1, the ␣5 and ␣6 subunits were acetylated with MAK3, and the ␤4 subunit was acetylated with NAT3. Furthermore, the Ac-MetPhe-Leu and Ac-Met-Phe-Arg termini of the ␣5 and ␣6 subunits, respectively, extended the known types of MAK3 substrates. Thus, nine subunits were N ␣ -acetylated, whereas the remaining five were processed, resulting in the loss of the N-terminal region. The 20 S proteasomes derived from either the nat1 mutant or the normal strain were similar in respect to chymotrypsinlike, trypsin-like, and peptidylglutamyl peptide hydrolyzing activities in vitro, suggesting that N ␣ -acetylation does not play a major functional role in these activities. However, the chymotrypsin-like activity in the absence of sodium dodecyl sulfate was slightly higher in the nat1 mutant than in the normal strain.

Fish, seafood and milk consumption seems to be beneficial, while butter, margarine, rapeseed oil, fast food and soft drinks could be risk factors for allergy and respiratory health.

Previously, we isolated a 4-kDa peptide capable of binding to a 43-kDa receptor-like protein and stimulating protein kinase activity of the 43-kDa protein in soybean. Both of them were found to localize in the plasma membranes and cell walls. Here, we report the physiological effects of 4-kDa peptide expressed transiently in the cultured carrot and bird's-foot trefoil cells transfected with pBI 121 plasmid containing the 4-kDa peptide gene. At early developmental stage, the transgenic callus grew rapidly compared to the wild callus in both species. Cell proliferation of in vitro cultured nonembryogenic carrot callus was apparently affected with the 4-kDa peptide in the medium. Complementary DNAs encoding the 4-kDa peptide from mung bean and azuki bean were cloned by PCR and sequenced. The amino-acid sequences deduced from the nucleotide sequences are homologous among legume species, particularly, the sites of cysteine residues are highly conserved. This conserved sequence reflects the importance of intradisulfide bonds required for the 4-kDa peptide to perform its function. Three dimensional structure of the 4-kDa peptide determined by NMR spectroscopy suggests that this peptide is a T-knot scaffold containing three b-strands, and the specific binding activity to the 43-kDa protein and stimulatory effect on the protein phosphorylation could be attributed to the spatial arrangements of hydrophobic residues at the solvent-exposed surface of two-stranded b-sheet of 4-kDa peptide. The importance of these residues for the 4-kDa peptide to bind to the 43-kDa protein was indicated by site-directed mutagenesis. These results suggest that the 4-kDa peptide is a hormone-like peptide and the 43-kDa protein is involved in cellular signal transduction of the peptide.Keywords: hormone-like peptide; plants; NMR; threedimensional structure; site-directed mutagenesis; physiological function.A 43-kDa protein found in the soybean seeds is a glycoprotein with sedimentation coefficient of 7S and isoelectric point ranging from 9.05 to 9.26 [1]. This protein has been classified into the category of globulin, which is soluble only in high ionic strength of salt solutions [1]. It consists of a and b subunits linked by disulfide bridge(s). There are a cysteine-rich domain in the N-terminal side of a subunit, a putative transmembrane domain in the b subunit [2], and a consensus sequence of ATP-binding site indispensable for protein phosphorylation activity [2]. The 43-kDa protein has autophosphorylation activity and protein kinase activity about two thirds of tyrosine kinase activity of the rat insulin receptor [3]. Immunocytochemistry has indicated that the 43-kDa protein is localized in the plasma membranes and the middle lamellae of cell walls [4], suggesting that it is a receptor-like protein. Western blotting and DNA cloning experiments revealed that these proteins are structurally similar to the 43-kDa protein and distribute in a number of legume species such as azuki bean, cowpea, French bean, lupin, mung bean and winged bean [5,6]...

Purpose:There is an increasing concern about sick building syndrome (SBS), especially in Asia. The aim of this study is to investigate associations between SBS and the home, school environment and personal factors among Japanese junior high school students.Methods:We investigated students in four junior high schools in Hyogo in Kansai area, Japan. A questionnaire study was performed among students (n=1056), 12-15 years old. Temperature and relative air humidity was measured in the classrooms and dust was collected from the classroom floors and air and was analysed for cat and dog allergens. Associations were analysed by multi-level logistic regression.Results:Mucosal symptoms (45.4%), general symptoms (38.9%) and skin symptoms (22.6%) were common. Totally 8.8% reported cat allergy, 6.1% dog allergy, 6.0% mold allergy and 25.7% pollen allergy. Atopy, window pane condensation, floor dampness and odor at home and high relative air humidity in the classrooms were associated with SBS.Conclusion:The prevalence of SBS symptoms was high and associated with both home and school environment. Window pane condensation and floor dampness at home can increase the risk for SBS symptoms in students. Moreover high relative air humidity at school may increase the risk for SBS.

Turmeric is widely used in Indian cuisine. The main constituents of turmeric are curcumin and its analogues, which are well-known antioxidant compounds. In the present study, we hypothesized that turmeric in curry might increase bowel motility and activate hydrogen-producing bacterial flora in the colon, thereby increasing the concentration of breath hydrogen. Eight healthy subjects fasted for 12 h and ingested curry and rice with or without turmeric (turmeric knockout curry). Breath-hydrogen concentrations were analyzed every 15 min for 6 h by gas chromatography with a semiconductor detector. Curry with turmeric significantly increased the area under the curve of breath hydrogen and shortened small-bowel transit time, compared with curry not containing turmeric. These results suggested that dietary turmeric activated bowel motility and carbohydrate colonic fermentation.

Starch granule-bound proteins in endosperms of common wheat (Triticum aestivum L.) included at least one major protein with a molecular weight of 61 000 (Wx protein:  starch granule-bound starch synthase) and six minor proteins with molecular weights of 115 000, 108 000, 100 000, 92 000, 80 000, and 15 000 (SGP-A1, -D1, -B1, -2, -3, and friabilin, respectively). Peptide mapping of SGP-A1, -D1, -B1, -2, and -3 indicated that the primary structures of SGP-A1, -D1, and -B1 are highly homologous but differ considerably from that of SGP-2. The internal sequences of peptides obtained from SGP-D1, -2, and -3 showed that SGP-D1 and SGP-3 are structurally similar to rice soluble starch synthase, whereas SGP-2 has homology to rice starch branching enzyme 3. These results are in agreement with results of immunological and enzymatic studies suggesting that SGP-A1, -D1, -B1, and -3 are types of soluble starch synthases and that SGP-2 is a starch branching enzyme. Keywords: Starch granule-bound protein; amino acid sequence; wheat (Triticum aestivum L.)

Using direct N-terminal analysis, only 31 N-terminally unblocked proteins out of 100 rice embryo proteins could be identified. To obtain protein sequence information for the remaining 69 blocked proteins, we developed a simple, efficient and rapid method. Using this method, we determined the peptide maps of 20 proteins per day in 10 pmol amounts. Applying this method to rice proteome analysis, we determined the internal sequences of all 69 blocked proteins. A total of 28 proteins out of 100 analyzed showed sequence similarity to the proteins with known functions in the SWISS-PROT and NCBI databases. Alternatively, we also used peptide mass fingerprinting determined by matrix assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) to identify the rice proteins separated by two-dimensional electrophoresis (2-DE). Although peptide-mass fingerprinting is a high-throughput method, we could not easily identify all the rice proteins or genes by this method, because the complete database information on rice, is not yet available and many proteins are post-translationally modified. Therefore, at present, the improved peptide mapping method as we report here is considered to be very useful in rice proteome analysis, especially for blocked proteins.

A study was undertaken to determine the association between the home environment along with the respiratory and allergy symptoms reported among female university students in Japan (N ¼ 153). A multiple logistic regression was applied for the determination and the study was controlled based on the age of the students, whether the student smokes and history of asthma/allergy suffering in parents. The characteristics of the population were: 15.7% are diagnosed asthma, 56.9% suffered pollen allergy, 15.7% cat allergy, 11.1% dog allergy, 25.0% wheeze in breathing, 24.2% suffered daytime breathlessness and 9.3% suffered nocturnal attacks of breathlessness. Evaluated characteristics were: 41.2% had a pet at home, 21.6% environmental tobacco smoke (ETS), 20.9% current dampness or moulds, 6.6% recent indoor painting, 7.2% new floor material, and 56.2% window condensation at home in winter. Wheeze was more common in concrete houses (OR ¼ 3.33; p50.05) and homes with water leakage and moulds infestation in the last 5 years (OR ¼ 3.75; p50.05), window condensation (OR ¼ 2.93; p50.05) and ETS (OR ¼ 3.04; p50.05). Nocturnal breathlessness was related to current building dampness (OR ¼ 7.48; p50.05), in particular to floor dampness. Furry pet allergy was more common in newer ( p50.05) and newly painted homes (OR ¼ 6.59; p50.05). Airway infections were more common in homes with new floor material (OR ¼ 4.19; p50.05). In conclusion, asthma, airway infections and allergies were common among female university students and related to home environmental exposures.