• International Journal of Industrial Entomology and Biomaterials
• /
• v.14 no.1
• /
• pp.37-44
• /
• 2007

A study was made of the haemolymph protein spectrum of mulberry silkworm (Bomhyx mori L.) from the first larval instar to imago. Horizontal starch gel electrophoresis was used. Sixteen races and eight F1 interracial hybrids, raised in Bulgaria, were analyzed. During the ontogenesis, a total of 17 protein bands (15 cathodic and 2 anodic) were detected. Distinct dynamics in the haemolymph protein spectrum was observed, in result of different expression during the individual development associated with the processes of growth, histolysis and histogenesis. Based on the ontogenetic dynamics found, a correspondence was assumed between some proteins detected by us using the starch gel electrophoresis and major haemolymph proteins (SP1, SP2, MHPs and Vg) detected by other authors using the polyacrilamide gel electrophoresis. Intraracial and interracial polymorphism was observed in four protein zones. The effect of four polymorphic loci with codominant and null alleles was suggested.

• Journal of Crop Science and Biotechnology
• /
• v.11 no.4
• /
• pp.227-232
• /
• 2008

Drought stress is one of the major abiotic stresses in agriculture worldwide. We report here a proteomic approach to investigate the impact of post-fertilization drought on grain quality in rice seed endosperm (Oryza sativa cv. IR-64). Plants were stressed for 4 days at 3 days before heading. Total proteins of endosperm were extracted and separated by two-dimensional gel electrophoresis. Not many protein spots showed differential accumulation in drought-stressed samples. More than 400 protein spots were reproducibly detected, including three that were up-regulated and five down-regulated. Mass spectrometry analysis and database searching helped us to identify six spots representing different proteins. Functionally, the identified proteins were related to protein synthesis and carbohydrate metabolism, such as Granule-Bound Starch Synthase (GBSS, Wx protein), which is thought to play a very important role in starch biosynthesis and quality, a very crucial factor in determining rice grain quality.

• The Korean Journal of Food And Nutrition
• /
• v.6 no.2
• /
• pp.73-80
• /
• 1993

In order to examine food components of coho salmon and rainbow-trout, We analyzed the composition of protein, amino acids and total lipids. The coho salmon muscle contained about 19.3% of protein with the composition of 29.9% in sarcoplasmic protein, 56.3oA in myofibrillar protein 12.5% alkali soluble protein and 2.6% in stroma. Those of rainbow-trout contained 34.1%, 56.4%, 8.3% and 2.9%, respectively. The sarcoplasmic and myofibrillar protein were composed of 13 subunits in coho salmon, and 16 and 15 subunits in rainbow-trout. Judging from the contents of essential amino acids, both muscle proteins were complete proteins. The most remarkable feature of free amino acids was that a large amount of dipeptide anserine was present with fairly lower levels of 1 methyl histidine, taurine, histidine, alanine and glycine in both muscle extracts. The total fatty acids of coho salmon was composed of 31.49% polyenes, 43.79% monoenes and 24.73% saturates. The composition of total fatty acid of coho salmon muscle was not different from that of rainbow-trout muscle.

• Journal of Plant Biotechnology
• /
• v.30 no.3
• /
• pp.281-291
• /
• 2003

In this review, we described the catalogues of the rice proteome which were constructed in our program, and functional characterization of some of these proteins was discussed. Mass-spectrometry is the most prevalent technique to rapidly identify a large number of proteome analysis. However, the conventional Western blotting/sequencing technique has been used in many laboratories. As a first step to efficiently construct protein cata-file in proteome analysis of major cereals, we have analyzed the N-terminal sequences of 100 rice embryo proteins and 70 wheat spike proteins separated by two-dimensional electrophoresis. Edman degradation revealed the N-terminal peptide sequences of only 31 rice proteins and 47 wheat proteins, suggesting that the rest of separated protein sports are N-terminally blocked. To efficiently determine the internal sequence of blocked proteins, we have developed a modified Cleveland peptide mapping method. Using this above method, the internal sequences of all blocked rice proteins(i, e., 69 proteins) were determined. Among these 100 rice proteins, thirty were proteins for which homologous sequence in the rice genome database could be identified. However, the rest of the proteins lacked homologous proteins. This appears to be consistent with the fact that about 45% of total rice cDNA have been deposited in the EMBL database. Also, the major proteins involved in the growth and development of rice can be identified using the proteome approach. Some of these proteins, including a calcium-binding protein that tuned out to be calreticulin, gibberellin-binding protein, which is ribulose-1.5-bisphosphate carboxylase/oxygense active in rice, and leginsulin-binding protein in soybean have functions in the signal transduction pathway. Proteomics is well suited not only to determine interaction between pairs of proteins, but also to identify multisubunit complexes. Currently, a protein-protein interaction database for plant proteins(http://genome.c.kanazawa-u.ac.jp/Y2H)could be a very useful tool for the plant research community. Also, the information thus obtained from the plant proteome would be helpful in predicting the function of the unknown proteins and would be useful be in the plant molecular breeding.

• Molecules and Cells
• /
• v.21 no.3
• /
• pp.381-388
• /
• 2006

Heat shock proteins (Hsp) 70 are a ubiquitous family of molecular chaperones involved in many cellular processes. A yeast strain, ssa1/2, with two functionally redundant cytosolic Hsp70s (SSA1 and SSA2) deleted shows thermotolerance comparable to mildly heatshocked wild type yeast, as well as increased protein synthesis and ubiquitin-proteasome protein degradation. Since mRNA abundance does not always correlate well with protein expression levels it is essential to study proteins directly. We used a gel-based approach to identify stress-responsive proteins in the ssa1/2 mutant and identified 43 differentially expressed spots. These were trypsin-digested and analyzed by nano electrospray ionization liquid chromatography tandem mass spectrometry (nESI-LC-MS/MS). A total of 22 non-redundant proteins were identified, 11 of which were confirmed by N-terminal sequencing. Nine proteins, most of which were up-regulated (2-fold or more) in the ssa1/2 mutant, proved to be stress-inducible proteins such as molecular chaperones and anti-oxidant proteins, or proteins related to carbohydrate metabolism. Interestingly, a translational factor Hyp2p up-regulated in the mutant was also found to be highly phosphorylated. These results indicate that the cytosolic Hsp70s, Ssa1p and Ssa2p, regulate an abundance of proteins mainly involved in stress responses and protein synthesis.

• Journal of Embryo Transfer
• /
• v.25 no.4
• /
• pp.281-286
• /
• 2010

This study was to evaluate the protein profile of seminal plasma using 2-DE in Hanwoo. Seminal plasma was harvested from five mature Hanwoo, and seminal plasma protein was extracted by M-PER Mammalian Protein Extraction Reagent. Proteins were refined by clean-up kit and quantified by Bradford method until total protein was $300\;{\mu}l$. Immobilized pH gradient (IPG) strip was used 18 cm and 3~11 NL. SDS-PAGE was used 12% acrylamide gel. Each gels were visualized by comassie brilliant blue and silver staining. These spots were analyzed by MALDI-TOF MS and searched on NCBInr. The result, 20 proteins of 36 protein spots were searched through peptide sequencing on the NCBInr. 8 proteins profiled by 2-DE were proved through previous bovine studies and the name of each protein was albumin, nucleobindin, clusterin, TIMP-2, spermadhesin Z13, spermadhesin-1 and BSP proteins (BSP 30 kDa and BSP A1/A2). 12 new proteins were ATP synthase, protein MAK16 homolog, Transmembrane protein 214, E3 ubiquitin-protein ligase BRE1A, dual serine/threonine and tyrosine protein kinase, tissue factor pathway inhibitor 2, alpha-actinin-4, RUN domain-containing protein 3B, catenin alpha-1, protein-glutamine gamma-glutamyltransferase 2, plakophilin-1 and inter-alpha-trypsin inhibitor heavy chain H1 has not been previously described in the bovine seminal plasma study. These proteins may be contribute to define the type of proteins affecting fertility of male and improve the fertilizing ability of semen in Hanwoo.

• Microbiology and Biotechnology Letters
• /
• v.18 no.5
• /
• pp.541-546
• /
• 1990

Bacillus sphaericus ts-Dl290 was characterized by SDS-PAGE produced by the mutant at $30^{\circ}C$ and $42^{\circ}C$. The total amount of proteins produced by the mutant at $42^{\circ}C$ decreased to one-fifth of those at $30^{\circ}C$; however, when the culture was shifted down from $42^{\circ}C$ after 4 to $30^{\circ}C$, the total amount of protein decreased to one-third and the 221 kd protein did not appear, but the 155 kd appeared remarkably. When the mutant and the wild type strain were cultured in the media containing 80$\mu g$ per ml of chloramphenicol at $42^{\circ}C$, the wild type strain synthesized half amounts of the total proteins than those at $30^{\circ}C$, and the mutant produced one-tenth of the total protein amounts. When the both strains were cultured in the media containing chloramphenicol, the 155 kd protein was produced was produced in lesser amounts than those without chloramphenicol. The 150 kd protein showed lethal activity to Culex pipiens 3rd instar larvae.

• Carbon letters
• /
• v.8 no.3
• /
• pp.184-190
• /
• 2007

Carbon Nano Tubes could be either metallic or semi-conducting in nature, depending on their diameter. Its photocatalytic behavior has given an impetus to use it as an anti-microbial agent. More than 95% Escherichia coli and Staphylococcus aureus bacteria got killed when exposed to Carbon Nano Tubes for 30 minutes in presence of sunlight. Carbon Nano Tubes are supposed to have smooth surface on to which it accumulates positive charges when exposed to light. The surface that is non illuminated has negative charge. At the cellular level microorganisms produce negative charges on the cell membrane, Therefore damaging effect of multi walled carbon nano tubes (exposed to light) on the microorganisms is possible. In this paper, photo catalytic killing of microbes by multi walled carbon nano tubes is reported. Killing was due to damage in the cell membrane, as seen in SEM micrographs. Moreover biochemical analysis of membrane as well as total cellular proteins by SDS PAGE showed that there was denaturation of membrane proteins as well as total proteins of both the microbes studied. The killed microbes that showed a decrease in number of protein bands (i.e. due to breaking down of proteins) also showed an increase in level of free amino acids in microbes. This further confirmed that proteins got denatured or broken down into shorter units of amino acids. Increased level of free amino acids was recorded in both the microbes treated with multi walled carbon nano tubes and sunlight.

• Journal of Ginseng Research
• /
• v.19 no.1
• /
• pp.39-44
• /
• 1995

Total saponin from Korean red ginseng changed thermodynamic parameters of membranes from dipalmitoylphosphatidylcholine (DPPC) and ghost erythrocytes of human. In liposomes from DPPC, temperature of the main transition (Lb'-La) in liquid-crystalline phase increases by 0.2$^{\circ}C$ in average, but enthalpy does not change. Total saponin at a concentration of smaller than $10^5$% "stabilizes" the timid bilayers. At larger than 0.07 of saponin/DPPC ratio, saponin leads to an exclusion of the bound lipid molecules from the main phase transition into lamella liquid crystalline La-phase. Total saponin influences specifically all erythrocyte membrane transitions in a concentration-dependent manner, i.e. on the structures of all the main membrane skeleton proteins. A high structural specificity of saponin with membrane proteins, could be a base of specificity of physiological response of not only erythrocytes, but also other cells.her cells.

• KOREAN JOURNAL OF CROP SCIENCE
• /
• v.51 no.spc1
• /
• pp.92-102
• /
• 2006

Abundant proteins often cause problems in proteome study. Glutelin family proteins (hereafter referred to glutelin) are present in rice proteome sample as over-whelming constituents with very high abundance. In order to increase the number of identified proteins in rice proteome study, we developed a newly improved method for sample preparation through the removal of glutelin. When the protein samples from rice seed were extracted by the conventional trichloroacetic acid (TCA) extraction method, glutelin accounts for about 60% of total rice seed proteins in SDS gels. Using our new water extraction method, glutelin consists of only about 10% of total proteins. After analyzing on a two-dimensional gel electrophoresis (2-DE), 937 protein spots were detected using the conventional TCA extraction method. On the other hand, 1240 proteins could be seen using the new water extraction method. The selectivity for non-glutelin and less abundant protein by the water extraction method was also confirmed by ESI-Q/TOF mass spectrometry analysis. Thus, the new water extraction method developed here can be efficiently used to study the proteome analysis of rice storage seed.