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Biomedical subjects

K Tao

Publications and source records attributed to K Tao.

At least 37 records · Page 2Linked to original sources
PubMed1997

oxyR-dependent induction of Escherichia coli grx gene expression by peroxide stress.

The Escherichia coli OxyR protein is a transcriptional activator for a number of genes induced in response to low concentrations of hydrogen peroxide. To identify additional OxyR-regulated genes, I cloned a DNA fragment that shows promoter activity regulated by OxyR by direct selection of OxyR-binding DNA fragments. Analyses of the cloned fragment indicate that the grx gene, encoding glutaredoxin 1, is inducible by hydrogen peroxide in an oxyR-dependent fashion.

Base Sequence↗
PubMed1997

[Diagnosis and treatment of congenital choledochal cyst].

From January 1980 to June 1996, 82 patients with congenital choledochal cysts were treated and 76 of them were operated on B-ultrasonic diagnosis was made with a correct diaginostic rate of 93.9%. The total effective rate was was 78.4% after follow-up for 84 months. Resection of the cyst with Roux-Y hepaticojejunostomy was successful in 96.2% of the patients. Outer drainage was used as the first-aid measure. Cystoduodenostomy and cystojejunostoy which may lead to complications should be abandoned.

Adolescent↗
PubMed1997

Expression in Escherichia coli and purification of human thrombopoietin.

Human thrombopoietin (TPO) has been successfully overexpressed in Escherichia coli, with an expression level of about 12% of total cellular protein. The full-length TPO gene was subcloned into the prokaryotic expression vector pKK233-2 under the control of the inducible tac promoter. The recombinant protein was produced mainly in the form of inclusion body. By efficient renaturation and one-step purification, the recombinant protein was purified to homogeneity. The specific activity and yield of recombinant TPO can reach 2 x 10(4) units/mg and 2 mg/g of wet E. coli cells respectively.

Chromatography, Liquid↗
PubMed1996

Mutagenicity of high fat diets in the colon and small intestine of transgenic mice.

Dietary fat has been implicated as a cause of colon cancer by epidemiological studies. Unfortunately, these studies are compatible with high fat as either initiator or promoter. Since initiators are normally mutagenic, we have tested the mutagenicity of high fat diets in the intestinal epithelium at two loci. The Dlb-1 assay, used in the small intestine, detects a wide spectrum of mutations. The lacI assay (Big Blue Mouse assay) is not as sensitive to some types of mutation as is Dlb-1 but was used in the colonic epithelium. Mice suitable for both assays were fed isocaloric high fat diets and subsequently assayed for somatic mutation. The diets consisted of: (i) a mixture of beef tallow, butter and lard totalling to 31% w/w of the diet (AIN-76A) up to 17 weeks; and (ii) corn oil, beef tallow, lard or butter individually, at 31% w/w of the diet for 5 and 9 weeks. These diets provided 50% of the calories from fat. The weights of the experimental and control mice were similar throughout the experiment. No significant increases in mutant frequencies were observed on any high fat diet compared to controls, so we conclude that uncooked fats are not mutagenic and are not initiators of carcinogenesis in the intestinal epithelium.

Animals↗
PubMed1995

Lysine-based structure in the proregion of procathepsin L is the recognition site for mannose phosphorylation.

The recognition of lysosomal enzymes by UDP-GlcNAc: lysosomal-enzyme GlcNAc-1-phosphotransferase (phosphotransferase) is mediated by a protein structure on lysosomal enzymes. It has been previously demonstrated that lysine residues are required for phosphorylation of procathepsin L and are a common feature of the site on many lysosomal proteins. In this work, the procathepsin L recognition structure was further defined by identification of the region of the protein containing the structure and the critical lysine residues involved. Removal of the cathepsin L propeptide by low pH-induced autocatalytic processing abolished phosphorylation. The addition of either the purified propeptide or a glutathione S-transferase-propeptide fusion protein to the processed protein restored phosphorylation. Mutagenesis of individual lysine residues demonstrated that two propeptide lysine residues (Lys-54 and Lys-99) were required for efficient phosphorylation of procathepsin L. By comparison of the phosphorylation rates of procathepsin L, lysine-modified procathepsin L, and the procathepsin L oligosaccharide, lysine residues were shown to account for most, if not all, of the protein-dependent interaction. On this basis, it is concluded that the proregion lysine residues are the major elements of the procathepsin L recognition site. In addition, lysine residues in cathepsin D were shown to be as important for phosphorylation as those in procathepsin L, supporting a general model of the recognition site as a specific three-dimensional arrangement of lysine residues exposed on the surface of lysosomal proteins.

3T3 Cells↗
PubMed1995

The transmission rate of the lacI transgene from the Big Blue mouse.

Since transgenic mice are being used to analyze somatic and germinal mutation rates in vivo, it is of interest to know to what extent these mice are normal or abnormal in any way. During experiments designed to compare the mutational response of the transgene and an endogenous gene, Big Blue mice hemizygous for the transgene were bred to create a hybrid mouse in which the comparisons could be made. The fraction of these mice that inherited the transgene was 37% rather than the Mendelian expectation of 50%.

Animals↗
PubMed1995

Mapping of the OxyR protein contact site in the C-terminal region of RNA polymerase alpha subunit.

The Escherichia coli OxyR protein requires the C-terminal contact site I region of the RNA polymerase alpha subunit for cooperative interaction with and transcription activation at OxyR-dependent promoters, suggesting direct protein-protein contact between OxyR and the C-terminal region of the alpha subunit. To determine the precise location of the OxyR protein contact site(s) in this region, we carried out mutational analysis of the 3' half of E. coli rpoA, the gene encoding the alpha subunit of RNA polymerase. We isolated a number of rpoA mutants defective in oxyR-dependent transcription activation at the E. coli katG promoter. Nucleotide sequence analysis of the rpoA gene from these mutants revealed that the mutations showing clear phenotypes are all clustered at two narrow regions (amino acid residues 265 to 269 and 293 to 300) within the C terminus of the alpha subunit. Reconstituted RNA polymerases containing the mutant alpha subunits were unable to respond to transcription activation in vitro at the katG, ahpC, and oxyX promoters by OxyR. These results suggest that these two regions comprise the contact surfaces on the alpha subunit for OxyR.

Bacterial Proteins↗
PubMed1994

The proregion of cathepsin L is required for proper folding, stability, and ER exit.

To investigate the role of the proregion in the biosynthesis and trafficking of mouse cathepsin L, cathepsin L cDNAs encoding proteins with altered proregions were constructed and their expression in COS cells was examined. As in transformed cells, normal mouse cathepsin L was secreted by COS cells. In contrast, two altered proregion cathepsin L proteins, one in which the proregion was deleted and a second in which the proregion was replaced with that of a homologous protein (aleurain), were retained within the cell and degraded over a period of 2-6 h. Immunofluorescence localization and the lack of effect of NH4Cl and brefeldin A on the turnover of the altered cathepsin L proteins indicated that their degradation occurred in the endoplasmic reticulum (ER). By using brefeldin A to induce colocalization of the UDPGlcNAc: lysosomal enzyme N-acetylglucosamine-1-phosphotransferase with the cathepsin L proteins in the ER, it was shown that the altered proteins were not susceptible to mannose phosphorylation as they exist in the ER. Trypsin sensitivity assays indicated that altered proregion proteins synthesized in COS cells or in vitro are misfolded. Taken together, these results indicate that the proregion plays an essential role in proper folding of cathepsin L. ER retention, decreased stability, and lack of mannose phosphorylation of the altered proteins are most likely secondary effects resulting from improper folding.

3T3 Cells↗
PubMed1993

Involvement of the RNA polymerase alpha subunit C-terminal region in co-operative interaction and transcriptional activation with OxyR protein.

The role of the alpha subunit of Escherichia coli RNA polymerase in transcription activation by the OxyR protein was investigated using in-vitro-reconstituted RNA polymerase containing alpha subunits carrying C-terminal truncations or an amino acid substitution. Mutant RNA polymerases failed to respond to transcription activation of the E. coli OxyR-dependent promoters. DNase I footprinting analysis indicates that the OxyR protein exerts a co-operative effect on the binding of wild-type RNA polymerase, but not the mutant RNA polymerases, to the katG promoter. Together, these results suggest that direct protein-protein contact between the OxyR protein and the C-terminal contact site I region of the RNA polymerase alpha subunit plays an essential role in transcription activation at the OxyR-dependent promoters.

Bacterial Proteins↗
PubMed1991

[IgA nephropathy in Chinese and Australian patients: a comparison between clinical and pathological features].

The clinical and pathological data were compared between 88 Chinese and 88 Australian patients with IgA nephropathy, whose age, sex and course of disease identified by renal biopsy were matched. Statistical differences showed: More Chinese patients had edema and loin pain, while more Australian patients had hypertension, glomerular sclerosis and arterial and/or arteriolar abnormalities; impairment of renal function correlated with crescent body formation and loin pain with severe hematuria were only found in Chinese patients, While correlations between severe hematuria and histological changes, hypertension and glomerular sclerosis, impaired renal function and glomerular sclerosis were only seen in Australian patients. Differences between the two groups in symptoms, histological changes and clinico-pathological correlations suggest that IgA nephropathy is a heterogeneous disease, it may result from more than one factor.

Adult↗
PubMed1991

Purification and characterization of the Escherichia coli OxyR protein, the positive regulator for a hydrogen peroxide-inducible regulon.

The Escherichia coli oxyR gene is required for the induction of a regulon that is inducible by hydrogen peroxide and confers resistance to oxidative stresses. We constructed a plasmid system that greatly overproduced OxyR protein and purified the protein. OxyR protein specifically bound to the upstream regulatory regions of the oxyR and katG genes as demonstrated by the gel-retardation assay and the DNase I footprinting experiment, and activated the transcription initiation of the katG gene in vitro. Using a plasmid carrying an oxyR'-'lacZ fusion gene, we studied the regulation of oxyR expression in vivo. The expression of oxyR was not induced by the treatment with a low concentration of hydrogen peroxide which induces the genes in the oxyR regulon. The expression of the oxyR'-'lacZ gene was higher in an oxyR-deletion strain than in the oxyR+ strain, and was repressed by overexpressing the OxyR protein. These results suggest that OxyR protein functions as a repressor for oxyR, in addition to its known function as a transcriptional activator for the genes in the oxyR regulon.

Bacterial Proteins↗
PubMed1989

Molecular cloning and nucleotide sequencing of oxyR, the positive regulatory gene of a regulon for an adaptive response to oxidative stress in Escherichia coli: homologies between OxyR protein and a family of bacterial activator proteins.

Treatment of Escherichia coli and Salmonella typhimurium cells with a low dose of hydrogen peroxide induces expression of a large number of genes, and confers resistance to oxidative stresses. The oxyR gene encodes a positive regulatory protein for a subset of these genes involved in the defense against oxidative damage. We cloned a DNA fragment that contains the E. coli oxyR region on a plasmid vector, and analyzed the nucleotide sequence of the gene. The amino acid sequence of OxyR protein, deduced from the nucleotide sequence, shows a high degree of homology to the sequences of a number of bacterial activator proteins including LysR, CysB, IlvY, MetR and NodD. The product of the oxyR gene identified by the maxicell procedure was a 34 kDa protein, which agrees with the size predicted from the nucleotide sequence of the gene.

Amino Acid Sequence↗
PubMed1989

Expression of complement 3 receptors (CR1 and CR3) on neutrophils and erythrocytes in patients with IgA nephropathy.

Expression of C3 receptors (CR1 and CR3) on neutrophils (PMN) was measured in 26 patients with IgA nephropathy (IgA N), 17 normal persons, and 8 patients with non-IgA glomerulonephritis (non-IgA GN) by fluorescence activated cell sorting after labeling with monoclonal antibodies. Mean channel fluorescence for both CR1 and CR3 on PMN was significantly higher in IgA N patients than in either control group (p less than 0.01). (CR1 mean +/- SD 42.5 +/- 10.4 in IgA N, 31.7 +/- 11.5 in normal controls, 27.8 +/- 9.0 in non-IgA GN; CR3 94.0 +/- 16.5, 75.0 +/- 16.6 and 76.7 +/- 15.6, respectively.) No differences were found between the two control groups or between IgA N patients with normal and impaired renal function. These results imply that PMN are activated in IgA N patients. The expression of CR1 and CR3 of PMN may be upregulated by immune complexes (ICs), enhancing both phagocytosis of C3b- or iC3b-coated particles, and absorptive pinocytosis of soluble ICs containing C3b or iC3b. Erythrocyte CR1 and CR3 expression was measured by ELISA and found to be slightly but significantly lower in IgA N patients than in the other 2 groups (CR1 85.3 +/- 28.4 in IgA N, 113.1 +/- 28.8 in normal controls, 109.4 +/- 28.2 in non-IgA GN, p less than 0.02; CR3 80.9 +/- 20.0, 100.4 +/- 19.9, 101.2 +/- 24.9, respectively, p less than 0.05).

Adult↗
PubMed1987

The roles of different excision-repair mechanisms in the resistance of Micrococcus luteus to UV and chemical mutagens.

M. luteus mutants showing increased sensitivity to both UV and 4-NQO were isolated after the treatment of parental ATCC4698 strain with MNNG. The mutants were also highly sensitive to mitomycin C, cis-platinum, 8-methoxypsoralen (8-MOP) plus near-UV and angelicin plus near-UV in various degrees. The endonuclease activity specific for pyrimidine dimers in UV-irradiated DNA was normally detected in extract of the mutants. With regard to host-cell reactivation ability the mutants fell into two groups. The hcr- mutants lacked the ability to reactivate UV-damaged N6 phage and were resistant to X-rays. The incision of DNA did not occur during incubation after the treatment with angelicin plus near-UV in the hcr- mutants, whereas it occurred in the parental strain. The facts indicate that the hcr- mutants are defective in the incision mechanism which has a wide substrate specificity, similar to the UVRABC nuclease of E. coli. On the other hand, the incision of DNA and the removal of UV-induced thymine dimers from DNA occurred in the hcr- mutants as well as in the parental strain, which is ascribed to the UV endonuclease activity. Compared with the hcr- mutants, hcr+ mutants were highly sensitive to X-rays, like recA- mutants of E. coli.

4-Nitroquinoline-1-oxide↗
PubMed1985

Isolation and purification of a small peptide with activity of increasing E-receptor expression from calf thymus.

A small peptide of molecular mass lower than 1,000 daltons was isolated and purified from a crude extract of calf thymus. It has been demonstrated to have the activity of increasing E-rosette formation and E-receptor expression of human and porcine T-lymphocytes with three different in vitro assays. Amino acid composition analysis showed that this peptide consists of glutamic acid, aspartic acid and glycine residues at a molecular ratio of 3:3:2. A hypothesis of a multi-factoral and multi-staged mechanism of regulation of thymocyte differentiation and maturation in the thymus is proposed.

Animals↗