PubMed Health⌕ Search

Biomedical subjects

C M Cheng

Publications and source records attributed to C M Cheng.

At least 37 records · Page 2Linked to original sources

Single-stranded DNA binding protein from bacteriophage cf: characterization, gene localization and protein-ssDNA complex.

The single-stranded DNA binding protein from the filamentous bacteriophage cf has been purified and characterized. The first 12 amino acids, resulting from the N-terminal amino acid sequencing analysis of the protein, agree with an open reading frame (ORF) on the cf genome. The ORF contains 294 bp and codes for a 98 a.a. protein of molecular weight 10.8 kDa, consistent with the result from the denaturing protein gel analysis. The protein appears to be a homodimer as evident from the apparent molecular weight of about 22 kDa obtained from native protein gel analysis. The gene location of the protein has been identified as gene V of the cf single stranded genome, same as that from the M13 phage. The GVP of cf shows a strong sequence homology to the ssDNA binding proteins of Ff, IKe and Pf3 filamentous phages. The DNA binding wing of GVP, conserved among the filamentous phages, has been predicted for cf. To further characterize the protein, the GVP-ssDNA complex of cf has been purified from the infected host (Xanthomonas campestris pv. citri) by density gradient centrifugation. Transmission electron microscopy (TEM) images of the complex showed that it is about 1200 nm in length and 9 nm in diameter and it has a highly regular morphology with a central groove shadow running along the entire structure, but without any apparent helical pattern seen in the M13 complex. The GVP-ssDNA complex of cf seems more rigid than that of M13. Our computer modeling study suggested that this difference between the two complexes may be due to the additional 11 or 12 amino acids at the C-terminal end of the cf-GVP.

Amino Acid Sequence↗

Rifampicin: an inhibitor of Xp12-specific protein phosphorylation in Xanthomonas oryzae pv. oryzae.

Phosphorylation of the three Xp12-specific phosphoproteins was drastically reduced by rifampicin, an antibiotic that specifically inhibits the host-cell RNA polymerase. However, this inhibitory effect could not be found in spontaneous mutants of Xanthomonas oryzae pv. oryzae whose RNA polymerase are resistant to the drug. The inhibitory effect of rifampicin treatment also resulted suppression of the Xp12 multiplication cycle. This implies the physiological significance of this effect and supports our previous prediction that phosphorylation plays an important role in the life cycle of Xp12. The acid- and alkali-labile character of the Xp12-specific phosphoproteins and the chemical stability of the phosphoryl linkages show that the corresponding protein kinase catalyzes the formation of an acyl phosphorylation. Subsequent fractionation of cell lysate revealed that the phosphoproteins were located in the periplasm. Actinomycin D, which affects transcription through DNA condensation rather than its binding to RNA polymerase, was not able to cause the inhibition effect. On the other hand, cerulenin was found to reduce the acyl phosphorylation which hints at a possible role of cell membrane in the phosphorylation. Here we present the evidence for the functional involvement of the rifampicin treatment on protein phosphorylation. A possible mechanism of rifampicin on the alternation of acyl phosphorylation is proposed.

Anti-Bacterial Agents↗

rpoS regulation of acid, heat, and salt tolerance in Escherichia coli O157:H7.

An rpoS mutant (rpoS::pRR10) of Escherichia coli O157:H7 ATCC 43895 was generated. Stationary-phase acid, heat, and salt tolerance was significantly reduced, and starvation-induced acid tolerance did not develop in the mutant. RpoS was also important for survival of E. coli O157:H7 in dry, fermented sausage.

Acids↗

Rapid inhibition of protein histidine phosphorylation by UV-irradiation in Xanthomonas oryzae pv. oryzae.

Exposure of Xanthomonas oryzae pv. oryzae cells to 254 nm UV radiation resulted in an alteration of protein phosphorylation. Labelling of the phosphohistidine-containing proteins with molecular masses of 81 and 32 kDa, named p81 and p32, was rapidly reduced following UV irradiation in the early exponential cells, but the decrease was not detected in mid-exponential cells. Mitomycin C, a DNA replication inhibitor, and rifampicin, a drug generally used to inhibit RNA synthesis and DNA replication, were also found to reduce the histidyl phosphorylation. However, this alteration of protein phosphorylation was not hindered by chloramphenicol treatment. A possible role for these histidyl phosphoproteins in sensing UV light is proposed.

Bacterial Proteins↗

Appearance of insulin-like growth factor mRNA in the liver and pyloric ceca of a teleost in response to exogenous growth hormone.

Augmentation of vertebrate growth by growth hormone (GH) is primarily due to its regulation of insulin-like growth factor I (IGF I) and IGF II levels. To characterize the effect of GH on the levels of IGF I and IGF II mRNA in a teleost, 10 micrograms of bovine GH (bGH) per g of body weight was administered to juvenile rainbow trout (Oncorhynchus mykiss) through i.p. injection. The levels of IGF I and IGF II mRNA were determined simultaneously, by using RNase protection assays, in the liver, pyloric ceca, kidney, and gill at 0, 1, 3, 6, 12, 24, 48, and 72 hr after injection. In the liver, IGF I mRNA levels were significantly elevated at 6 and 12 hr (approximately 2- to 3-fold, P < or = 0.01), while IGF II mRNA levels were significantly elevated at 3 and 6 hr (approximately 3-fold, P < or = 0.01). In the pyloric ceca, IGF II mRNA levels were significantly elevated at 12, 24, and 48 hr (approximately 3-fold, P < or = 0.01), while IGF I mRNA was below the limits of assay accuracy. GH-dependent IGF mRNA appearance was not detected in the gill and kidney. Serum bGH levels, determined by using a radioimmunoassay, were significantly elevated at 3 and 6 hr (P < 0.005). In primary hepatocyte culture, IGF I and IGF II mRNA levels increased in a bGH dose-dependent fashion, with ED50 values of approximately 45 and approximately 6 ng of bGH per ml, respectively. The GH-dependent appearance of IGF II mRNA in the liver and pyloric ceca suggests important roles for this peptide hormone exclusive of IGF I.

Animals↗

Production of a biologically active recombinant teleostean growth hormone in E. coli cells.

We have isolated and characterized several recombinant lambda phage clones carrying growth hormone (GH) cDNA of striped bass (Morone saxatilis). Nucleotide sequence and the predicted amino acid sequence of sbGH was determined from a recombinant clone carrying the longest cDNA insert. The sbGH cDNA encodes a pre-hormone of 204 amino acid residues. Comparison of the predicted amino acid sequence of sbGH with those of other vertebrates revealed different degrees of sequence identity: approximately 98% with European sea bass; 90% with bluefin tuna; bonito and red seabream; 71% with winter flounder; 64% with salmonids; 55% with carp; and 38% with human. Expression of the mature sbGH cDNA (without the signal peptide sequence) in E. coli cells under regulation of the lambda phage PL promoter produced a polypeptide of 20 kDa. Following renaturation, this recombinant hormone was shown to be biologically active in a radioreceptor competition binding assay and in the induction of hepatic insulin-like growth factor I (IGF-I) mRNA synthesis in vivo.

Amino Acid Sequence↗

Synergism of GH and IGF-I in stimulation of sulphate uptake by teleostean branchial cartilage in vitro.

The combined effects of teleostean GH and human IGF-I in the regulation of teleost branchial cartilage growth were examined. Ceratobranchial cartilages were dissected from the common carp (Cyprinus carpio) and maintained in a defined culture medium supplemented with recombinant striped bass GH (sbGH), human IFG-I (hIGF-I) or both, and the uptake of [35S]sulphate by cartilage explants was measured. sbGH alone in the culture medium did not exhibit a significant stimulatory effect on the uptake of [35S]sulphate when compared with the controls. However, with hIGF-I in the culture medium, as low as 1 ng/ml, the stimulatory effect of sbGH was apparent and dose-dependent. The synergism of sbGH and hIGF-I was observed at the concentrations of 1 and 10 ng IGF-I/ml tested. At a constant hIGF-I concentration (10 ng/ml), a maximum stimulatory effect was detected with 3 micrograms recombinant sbGH/ml, at which point a 2.3-fold increase in sulphation activity was obtained when compared with the treatment with hIGF-I alone. A similar dose-dependent stimulatory effect was observed when native common carp and bonito GHs were tested using this assay system. These results suggest that the synergistic effect of sbGH and hIGF-I on sulphation activity is a common biological function of teleost GH, thus ruling out the possibility of experimental artifacts resulting from utilization of heterologous GH. Furthermore, experiments were conducted to test whether the synergism between sbGH and hIGF-I occurred between sbGH and insulin.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Specific protein phosphorylation induced in Xanthomonas campestris pv. oryzae by bacteriophage Xp12.

We have investigated the endogenous phosphorylation patterns of phosphorylated proteins of Xanthomonas campestris pv. oryzae induced by its bacteriophages. For bacteriophage Xp12-infected cells, at least three phosphoproteins with apparent molecular weights of 28, 28.5 and 45 kDa were detected by in vitro labeling with [gamma-32P]-ATP. These Xp12-specific phosphoproteins only occurred with Xp12 infection, and were not shown in uninfected or Xp10-infected cells. The protein kinase(s) responsible could use either ATP or GTP as the nucleotide substrate with nearly the same efficiency. Magnesium was proved to be an essential factor for the phosphorylation. EGTA treatment excluding the possibility that the presumed protein kinase was calcium-dependent. Under our reaction conditions, the optimal phosphorylation occurred at pH 7 to 8, for 30 to 40 min at 25 to 37 degrees C. The Xp12-specific protein phosphorylation hint the existence of a physiological regulation mechanism involved in the life cycle of bacteriophage Xp12. Furthermore, the presumed protein kinase was shown to be encoded by the genome of Xp12 rather than indirectly induced by Xp12 infection.

Adenosine Triphosphate↗

Antitumor agents, 145. Cytotoxic asprellic acids A and C and asprellic acid B. new p-coumaroyl triterpenes, from Ilex asprella.

Three new p-coumaroyl triterpenes, asprellic acids A [1], B [2], and C [3], were isolated from the EtOH extract of dried leaves of Ilex asprella. The structures for 1-3 have been established as 3,27-di-O-trans-p-coumaroyl [1], 3-O-trans-p-coumaroyl-27-O-cis-p-coumaroyl- [2], and 3-O-cis-p-coumaroyl-27-O-trans-p-coumaroyl- [3] 3 beta,27-dihydroxyolean-12-en-28-oic acid, respectively, by spectral and chemical examinations. Asprellic acid A [1] exhibited potent cytotoxicity against the RPMI-7951 cell line with an ED50 value of 0.62 micrograms/ml, while the cytotoxicity of asprellic acid C [3] against RPMI-7951 was marginal (ED50 5.5 micrograms/ml). Compounds 1 and 3 also showed cytotoxicity against KB cells with ED50 values of 3.75 and 2.86 micrograms/ml, respectively. Asprellic acid B [2] did not show cytotoxicity (> 10 micrograms/ml) against KB or RPMI-7951 cell lines.

Adenocarcinoma↗

Characterization of the pp70 protein phosphorylation in the extract of rice young panicle.

The endogenous phosphorylation pattern of the extract prepared from rice young panicle has been examined. The extract was phosphorylated in vitro by incubating with [gamma-32p] ATP, and then analyzed by SDS-PAGE and autoradiography. At least eight major phosphoproteins with apparent molecular weights of 70, 60, 52, 40, 33, 30, 20, 16 and 15 kd were detected in the crude extract of rice young panicle. The pp70 which represents the major phosphoprotein in the crude extract of young panicle of rice is a cytosolic protein. Photoaffinity labeling with 8-azido-ATP revealed that a major protein with molecular weight 42,000 dalton but not pp70 can be specifically bound ATP. This suggests that pp70 is not a protein kinase itself, but a substrate of protein kinase. The in vitro phosphorylation of the pp70 occurs at a serine or threonine residue and is time and temperature dependent. The phosphorylation of pp70 does not depend by exogenous Ca++ or cAMP, suggesting that pp70 is a major substrate of an Ca++ or cAMP independent protein kinase in rice young panicle.

Calcium↗

[Combined methods of dialysis, cooked meat medium enrichment and laboratory animal toxicity for screening Clostridium botulinum spores in honey and infant food].

One hundred and fifty two samples of honey were purchased from apiaries and markets in Taiwan, meanwhile fifty samples of infant food including powders of infant milk, wheat, rice and commercial mixed cereals, as well as juice had been bought from supermarkets in Taipei city from July 1988 through April 1989. The samples were used for detecting the presence of Clostridium botulinum spores. Honey samples were prepared by dialysis to obtain the bacterial spores; however, infant food had been innoculated directly into the cooked meat medium. The suspicious isolated colonies were identified by Vitek automatic microbial identification system. The supernatants of cooked meat medium enrichment were performed for typing botulinum toxins via the laboratory animal toxicity test. The results showed that two samples of honey contained the spores of C. botulinum type B, but none in samples of infant food. The specific response of clinical signs in mice, after being administered the supernatant of suspected cultural broth, were also observed and described in this article.

Animals↗

Lateralization in the visual perception of Chinese characters and words.

It has been reported that tachistoscopic perception of single Chinese characters is better with a left-visual-field (LVF) than with a right-visual-field (RVF) presentation and that of Chinese words consisting of characters is better with a RVF presentation (O. J. L. Tzeng, D. L. Hung, B. Cotton, & S.-Y. Wang, 1979, Nature (London), 382, 499-501). In this study, the nature of this character-word difference in lateralization was explored in a task in which stimuli were presented unilaterally to a visual field for recognition test. Four types of stimuli were used: Single character, single pseudo- or noncharacter, two-character word, and two-character pseudoword. Results show (a) no visual-field advantage for illegal characters and words, (b) a LVF-advantage effect for characters associated with a more prominent LVF than RVF character-superiority effect, (c) a RVF-advantage effect for words associated with a more prominent RVF than LVF word-superiority effect, and (d) these two visual-field effects for characters and words being not absolute, they occur only with a low rather than with a high recognition for their respective illegal counterparts. These results suggest that the character-word difference is due to a more efficient lexical interpretation of character stimuli in the right than in the left hemisphere and a more efficient lexical interpretation of word stimuli in the left than in the right hemisphere.

Adult↗

Unusual origins of the buccal and mylohyoid nerves.

Dissection of the mandibular nerve in a cadaver of southern Chinese origin showed the mylohyoid nerve arising from the lingual nerve and the buccal nerve arising from the inferior alveolar nerve within the mandibular ramus. It is estimated that this variation in the origin of the buccal nerve occurs in 6.1% of the southern Chinese population.

Cheek↗

Affinity isolation and characterization of immunoglobulin G Fc fragment-binding glycoprotein from human blood platelets.

Immune complexes bind to several eukaryotic cell types including human blood platelets through the Fc fragment of immunoglobulin (Ig) G. Utilizing immobilized Fc fragment of IgG enabled us to isolate from human blood platelets a glycoprotein of an apparent Mr = 255,000 which, upon reduction, dissociated into sub-units of an apparent Mr = 50,000. This Fc fragment-binding glycoprotein has an isoelectric point between pH 6.3 and 6.9 and is composed of 34% hydrophobic, 25% acidic, and 14% basic amino acids. The Fc fragment-binding glycoprotein was also isolated from human platelet membrane preparations and was unaffected by prior treatment of platelets with thrombin. Isolated Fc fragment-binding glycoprotein formed an in vitro complex with aggregated immunoglobulin G. These results suggest that the isolated Fc fragment-binding component may prove useful in studies concerning the functional role of glycoproteins as cellular receptors for the Fc fragment of IgG.

Antigen-Antibody Complex↗