• Biomedical Science Letters
• /
• v.4 no.1
• /
• pp.73-76
• /
• 1998

The authors characterized the antigen proteins and some specific antibodies from Echinostoma hortense. Crude antigen extracted from E. hortense worm was analyzed by SDS-PAGE of the crude antigen showed 46 profiles between 200.2 - 8.2kDa, among which 200.2, 107.9, 86.8, 75, 69.8, 46.8, 43.5, 34.5, 20.9, 13.6, 12.6, 11.7, and 8.2kDa, protein profiles were strong. EITB resolved the specific IgG antibody into 17 profiles between 193 - 13.7kDa, among which 198, 123.4, 100.8, 91.1, 88.1, 62.8, 34.2, 32, 29.9, 18, 15.7, 13.7kDa profiles showed strong immunostain.

• Food Science of Animal Resources
• /
• v.25 no.2
• /
• pp.232-237
• /
• 2005

Lactoferrin is a member of the transferrin family of iron-binding glycoproteins. It is originally found in milk. In addition to its antibacterial and antiviral activities, lactoferrin has many other biological functions include anti-inflammatory properties, antitumor, cell growth-promoting activity as well as antioxidant effect In the present study, we report the production of recombinant bovine lactoferrin and lactoferrin N-lobe in the Rhodococcus erythropolis (R erythropolis) using pTip vector. The expression level was investigated in various range of temperature, and we could successfully expressed the bovine lactoferrin and lactoferrin N-lobe in R erythropolis at low temperature. The recombinant proteins were purified by Nickel-Nitrolotriacetic acid (Ni-NTA). The purified proteins were confirmed by SDS-PAGE and Western blot, which indicating that the recombinant proteins have a molecular weight of 80kDa and 43kDa for bovine lactoferrin and lactoferrin N-lobe, respectively.

• Applied Biological Chemistry
• /
• v.43 no.4
• /
• pp.231-235
• /
• 2000

The changes of calmodulin levels, calmodulin-binding proteins, and $Ca^{2+}$/calmodulin-dependent glutamate decarboxylase during the growth of tobacco suspension cells were investigated. Tobacco cells exhibited a typical growth curve, including an exponential growth phase between 3 and 5 days after inoculation, and an apparent stationary phase occurring after 5 day. Although slight changes were observed from sample to sample, calmodulin protein levels remained similar during the phases of culture growth. Several $Ca^{2+}-dependent$ calmodulin-binding proteins including 56, 46, 36, and 32-kDa proteins were detected in tobacco cell extracts. The 56-kDa protein was identified as glutamate decarboxylase by Western-blot analysis using an anti-GAD monoclonal antibody. The levels of GAD protein and the specific activity of GAD enzyme were highest during the middle exponential phase of the culture growth cycle. These data suggest that $Ca^{2+}$/calmodulin-dependent glutamate decarboxylase is modulated during the growth of tobacco suspension cells.

• Journal of Microbiology
• /
• v.43 no.1
• /
• pp.34-37
• /
• 2005

In the present study, the xylA gene encoding a thermostable xylose (glucose) isomerase was cloned from Streptomyces chibaensis J-59. The open reading frame of xylA (1167 bp) encoded a protein of 388 amino acids with a calculated molecular mass of about 43 kDa. The XylA showed high sequence homology (92% identity) with that of S. olivochromogenes. The xylose (glucose) isomerase was expressed in Escherichia coli and purified. The purified recombinant XylA had an apparent molecular mass of 45 kDa, which corresponds to the molecular mass calculated from the deduced amino acid and that of the purified wild-type enzyme. The N-terminal sequences (14 amino acid residues) of the purified protein revealed that the sequences were identical to that deduced from the DNA sequence of the xylA gene. The optimum temperature of the purified enzyme was $85^{\circ}C$ and the enzyme exhibited a high level of heat stability.

• Journal of Microbiology
• /
• v.39 no.3
• /
• pp.162-167
• /
• 2001

Methylovorus sp. strain SS1 DSM 11726 was found to grow continuously when it was transferred from 30$\^{C}$ to 40$\^{C}$ and 43$\^{C}$. A shift in growth temperature from 30$\^{C}$ to 45$\^{C}$, 47$\^{C}$ and 50$\^{C}$ reduced the viability of the cell population by more than 10$^2$, 10$^3$and 10$\^$5/ folds, respectively, after 1h cultivation. Cells transferred to 47$\^{C}$ and 50$\^{C}$ after preincubation for 15 min at 43$\^{C}$, however, exhibited 10-fold increase in viability. It was found that incubation for 15 min at 40$\^{C}$ of Methylovorus sp. strain SSl grown at 30$\^{C}$ was sufficient to accelerate the synthesis of a specific subset of proteins. The major heat shock proteins had apparent molecular masses of 90, 70, 66, 60, and 58 kDA. The 60 and 58 kDa proteins were found to cross-react with the antiserum raised against GroEL protein. The heat shock response persisted for over 1h. The shock proteins were stable for 90 min in the cell. Exposure of the cells to methanol induced proteins identical to the heat shock proteins. Addition of ethanol induced a unique protein with a molecular mass of about 40 kDa in addition to the heat-induced proteins. The proteins induced in paraquat-treated cells were different from the heat shock proteins, except the 70 and 60 kDa proteins.

• The Korean Journal of Zoology
• /
• v.33 no.1
• /
• pp.108-118
• /
• 1990

Fibronectin is a glycoprotein found in the extracellular matrix as well as in the serum, and has been known to exert pronouned effed on the myoblast fusion. Our previous studies have suggested that the decrease of fibronectin levels during myogenesis is due to the decreased availability of the receptor for the 28 kDa fragrnent of fibronetin. In the fusion-blocked myoblasts by EGTA, the levels of fibronetin and binding of 28 kDa fragment decreased but far less than the control level. In contrast, the levels of fibronetin and binding of 28 kDa fragment decreased to the control level in the myoblast released from the fusion block. On this account, we suggest that the decrease of fibronetin levels during myoblast fusion is closely associated with the loss or alteration of the receptor for 28 kDa fragment. Mild trypsin treatment decreased the binding of the 28 kDa fragment to the myoblasts significandy. Similarly, the presence of gangliosides in the binding media decreased the binding of the 28 kDa fragment in a dose-dependent manner. Furthermore, gel overlay of 125 I-28 kDa fragment on the SDS-PAGE of the myoblast homogenates revealed that the 28 kDa fragment bound to a 43 kDa protein and to gangliosides as well. These results suggest that myoblast fusion is correlated with decrease of the receptor for the 28 kDa fragment and that the receptor might be a glycoprotein that contains glyco-conjugate found in gangliosides.

• The Korean Journal of Mycology
• /
• v.40 no.2
• /
• pp.109-113
• /
• 2012

We have isolated an anti-complementary polysaccharide from the hot water extracts of Phellinus linteus mycelia. Anti-complementary polysaccharide, PL-5-IIIa, was purified by ultrafiltration, gel permeation chromatography using Sepharose CL-4B. GPC (Sepharose CL-4B) and its homogenicity was demonstrated by HPLC. Using gel permeation chromatography with standard dextrans, its molecular weight was determined as about 800,000 dalton. The purified PL-5-IIIa was identified as a protein bound polysaccharide comprising of 29.6% protein and 64.2% carbohydrate which was composed of fucose(15.8%), galactose(43.1%) and mannose(40.6%).

• Reproductive and Developmental Biology
• /
• v.29 no.3
• /
• pp.187-191
• /
• 2005

This study was designed to investigate the changes of quantity and quality of proteins in medium and cytoplasm during in vitro maturation of bovine oocytes. The total quantity of proteins in medium decreased from 0 to 4.5 hr, but increased from 13.5 to 18 hr after the onset of in vitro maturation. The total quantity of protein in cytoplasm increased from 0 to 4.5 hr, decreased from 4.5 to 9 hr, and increased after 18 hr after the onset of in vitro maturation. A total of 298 protein spots was detected on a gel of 2D SDS-PAGE form maturation medium. Among 28 protein spots expressed significant differences in their quantity, 8 proteins were identified by peptide mass fingerprinting (aldose reductase, alpha enolase, apolipoprotein A-1 precursor, 43kDa collectin precursor, heat shock 27kDa protein, plasminogen activator inhibitor-1 precursor, thrombospondin 1, transitional endoplasmic reticulum ATPase). Among total of 35 protein spots detected on gel of 2D SDS-PACE from oorytes cytoplasm, $\beta$-tubulin was identified by peptide mass fingerprinting.

• Animal cells and systems
• /
• v.3 no.2
• /
• pp.221-228
• /
• 1999

In order to find a biochemical marker for late Iysosomes, we characterized two cDNAs which were cloned by using a monoclonal antibody (mAb) against Iysosomes in Amoeba proteus as a probe. The two cDNAs, a 1.3-kb cDNA in pBSK-Iys45 and a 1.6-kb cDNA in pBSK-Iys60, were found to encode proteins homologous to pepstatin-insensitive carboxyl proteinases (PICPs). E. coli transformed with pBSK-Iys45 produced two immunopositive polypeptides (45 and 43 kDa) and the cDNA in 1274 bases encoded a 44,733-Da protein (Lys45) of 420 amino acids containing one site for a core oligosaccharide. On the other hand, E. coli transformed with pBSK-Iys60 produced several polypeptides (64, 54, 45, 41, and 37 kDa) reacting with the mAb. The cDNA contained 1629 bases and encoded a 59,231-Da protein (Lys60) of 530 amino acids containing two sites for asparagine-linked core oligosaccharides. These two cDNAs showed identities of 60.3% in nucleotide sequences and 23.6% in amino acid sequences. Lys45 and Lys60 appeared to share XXEFQK as a common antigenic domain. The amino acid sequence of the Lys45 protein showed 17.4% identity and 40.9% similarity to that of PICP from Pseudomonas sp. 101. On the other hand, Lys60 showed a 24.3% identity and 51.9% similarity with human Iysosomal PICP in the amino acid sequence. A putative active center for serine protease, GTS＊xxxxxFxG, was found to be conserved among PICP homologues. The two PICPs are the first reported enzymatic markers for late Iysosomes.

• BMB Reports
• /
• v.32 no.4
• /
• pp.363-369
• /
• 1999

The orf8(chlD) gene cloned from Streptomyces antibioticus T$\"{u}$99 was overexpressed using an E. coli system to confirm its biological function. Induction of the E. coli strain transformed with recombinant plasmid pRFJ 1031 containing orf8 resulted in the production of a 43,000 dalton protein. Glucose-1-phosphate thymidylyltransferase activity of the cell extract obtained from the transformed strain was 4-5 times higher than that of the control strain. The expressed protein was purified 18-fold from E. coli cell lysate using three chromatographic steps with a 17% overall recovery to near homogeneity. The N-terminal amino acid sequence of the purified protein agrees with the nucleotide sequence predicted from the orf8 gene. The SDS-PAGE estimated subunit mass of 43,000 dalton agrees well with that calculated from the amino acid composition deduced from the nucleotide sequence of the orf8 gene (43,000 Da). Also, the native enzyme has a monomeric structure with a molecular mass of 43,000 dalton. The purified protein showed glucose-1-phosphate thymidylyltransferase activity catalyzing a reversible bimolecular group transfer reaction, and was highly specific for dTTP and ${\alpha}$-D-glucose 1-phosphate as substrates in the forward reaction, and for dTDP-D-glucose and pyrophosphate in the reverse reaction.