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Y Tang

Publications and source records attributed to Y Tang.

At least 181 records · Page 10Linked to original sources

[IR spectra and structures of two trinuclear molybdenum cluster compounds coordinated by binary carboxylates].

Two trinuclear molybdenum cluster compounds coordinated by binary carboxylates compounds Mo3S4 (DTP)3(phthalate) (Py)(A) and Mo3S4 (DTP)3(malonate)(Py)(B) have been synthesized. IR absorption peaks of Mo3S4 core and the ligands are assigned with reference to those of Mo3S4(DTP)4(Py)(C). The assignment of stretching vibrations of carboxylate (COO-) for both (A) and (B) reveals much information corresponding to their molecular structures.

English Abstract↗

Biochemical and spectroscopic characterization of Escherichia coli aconitases (AcnA and AcnB).

Escherichia coli contains two major aconitases (Acns), AcnA and AcnB. They are distantly related monomeric Fe-S proteins that contain different arrangements of four structural domains. On the basis of the differential expression of the acnA and acnB genes, AcnA has been designated as an aerobic-stationary-phase enzyme that is specifically induced by iron and oxidative stress, whereas AcnB functions as the major citric-acid-cycle enzyme during exponential growth. The biochemical and kinetic properties of the purified enzymes have now shown that AcnA is more stable than AcnB, has a higher affinity for citrate, and operates optimally over a wider pH range, consistent with its role as a maintenance or survival enzyme during nutritional or oxidative stress. In contrast, the better performance at high substrate concentrations and greater instability of AcnB indicate that AcnB is specifically adapted to function as the main catabolic enzyme and, by inactivation, to rapidly modulate energy metabolism in response to oxidative or pH stress, either directly or indirectly by regulating post-transcriptional gene expression. EPR and magnetic-CD spectroscopy showed that the iron-sulphur clusters of the bacterial Acns (and their binding sites) strongly resemble those of the mammalian enzymes. The EPR and MCD spectra of the oxidized inactive form of AcnB confirmed the presence of a [3Fe-4S](1+) (S=1/2) cluster. Comparisons showed that the EPR spectrum of AcnB more closely resembled that of mammalian mitochondrial Acn (m-Acn), whereas the spectrum of AcnA more closely resembled that of the cytoplasmic enzyme (c-Acn). The MCD spectra revealed spectroscopic signatures similar to that of m-Acn. Reconstitution of the active [4Fe-4S](2+) forms followed by one-electron reduction gave rise to EPR spectra that are almost identical with those reported for the mammalian enzymes.

Aconitate Hydratase↗

Spinal and brain circuits to motoneurons of the bulbospongiosus muscle: retrograde transneuronal tracing with rabies virus.

Retrograde transneuronal tracing with rabies virus from the left bulbospongiosus muscle (BS) was used to identify the neural circuits underlying its peripheral and central activation. Rats were killed at 2, 3, 4, and 5 days post-inoculation (p.i.). Rabies immunolabelling was combined with immunohistochemical detection of choline acetyltransferase and oxytocin. Virus uptake was restricted to ipsilateral BS motoneurons (2 days p.i.). The onset of transfer (3 days p.i.) visualized interneurons in the dorsal grey commissure (DGC), intermediate zone, and sacral parasympathetic nucleus (SPN), mainly in DGC at L5-S1, and revealed synaptic connections between BS and external urethral sphincter motoneurons. At 4 and 5 days p.i., higher-order interneurons were labelled in other spinal areas and segments. Supraspinal labelling initially involved only Barrington's nucleus, nucleus reticularis magnocellularis, and paragigantocellularis lateralis (4 days p.i.). Later, labelling extended to other populations traditionally associated with control of sexual activity and micturition (periaqueductal grey, paraventricular nucleus, medial preoptic area, prefrontal cortex), but also indicated the intervention of somatic descending motor pathways (vestibulospinal and reticulospinal neurons, "hindlimb" regions of sensorimotor cortex and red nucleus) and cerebellar nuclei in multisynaptic innervation of the labelled motoneurons. Dual color immunofluorescence disclosed multisynaptic links between these motoneurons and thoracolumbar medial sympathetic (choline acetyltransferase-immunoreactive) neurons. In contrast, preganglionic neurons in SPN and most oxytocinergic neurons in paraventricular hypothalamic nucleus remained unlabelled, suggesting that parasympathetic and somatic outflow to pelvic organs are probably controlled by separate interneuronal populations and that oxytocinergic spinal projections are more likely to influence sacral autonomic rather than somatic outflow.

Animals↗

Study of the Dissolution of the Barium Sulfate (001) Surface with Hydrochloric Acid by Atomic Force Microscopy.

Noncontact atomic force microscopy (NC-AFM) has been used to investigate the morphological changes of a freshly cleaved (001) surface of barium sulfate (barite) etched with an aqueous solution of 0.1 M HCl at room temperature. Shallow triangular etch pits with a height of 3.6 Å were developed in atomically flat (001) terraces. The etching of the surface was found to proceed in a layer-by-layer dissolution process. Because the crystal structure of barite exhibits a two-fold screw axis parallel to the c axis, "alternating" etch pits were formed, with any two consecutive etch pits pointing opposite to each other. These etch pits became deeper and more elongated along the b axis with time. Copyright 1999 Academic Press.

Journal Article↗

Identification of the endophilins (SH3p4/p8/p13) as novel binding partners for the beta1-adrenergic receptor.

Several G-protein coupled receptors, such as the beta1-adrenergic receptor (beta1-AR), contain polyproline motifs within their intracellular domains. Such motifs in other proteins are known to mediate protein-protein interactions such as with Src homology (SH)3 domains. Accordingly, we used the proline-rich third intracellular loop of the beta1-AR either as a glutathione S-transferase fusion protein in biochemical "pull-down" assays or as bait in the yeast two-hybrid system to search for interacting proteins. Both approaches identified SH3p4/p8/p13 (also referred to as endophilin 1/2/3), a SH3 domain-containing protein family, as binding partners for the beta1-AR. In vitro and in human embryonic kidney (HEK) 293 cells, SH3p4 specifically binds to the third intracellular loop of the beta1-AR but not to that of the beta2-AR. Moreover, this interaction is mediated by the C-terminal SH3 domain of SH3p4. Functionally, overexpression of SH3p4 promotes agonist-induced internalization and modestly decreases the Gs coupling efficacy of beta1-ARs in HEK293 cells while having no effect on beta2-ARs. Thus, our studies demonstrate a role of the SH3p4/p8/p13 protein family in beta1-AR signaling and suggest that interaction between proline-rich motifs and SH3-containing proteins may represent a previously underappreciated aspect of G-protein coupled receptor signaling.

Adaptor Proteins, Signal Transducing↗

Identification of a new member of the tryptase family of mouse and human mast cell proteases which possesses a novel COOH-terminal hydrophobic extension.

Mapping of the tryptase locus on chromosome 17 revealed a novel gene 2.3 kilobase 3' of the mouse mast cell protease (mMCP) 6 gene. This 3.7-kilobase gene encodes the first example of a protease in the tryptase family that contains a membrane-spanning segment located at its COOH terminus. Comparative structural studies indicated that the putative transmembrane tryptase (TMT) possesses a unique substrate-binding cleft. As assessed by RNA blot analyses, mTMT is expressed in mice in both strain- and tissue-dependent manners. Thus, different transcriptional and/or post-transcriptional mechanisms are used to control the expression of mTMT in vivo. Analysis of the corresponding tryptase locus in the human genome resulted in the isolation and characterization of the hTMT gene. The hTMT transcript is expressed in numerous tissues and is also translated. Analysis of the tryptase family of genes in mice and humans now indicates that a primordial serine protease gene duplicated early and often during the evolution of mammals to generate a panel of homologous tryptases in each species that differ in their tissue expression, substrate specificities, and physical properties.

Amino Acid Sequence↗

Use of a peptide mimotope to guide the humanization of MRK-16, an anti-P-glycoprotein monoclonal antibody.

A mimotope-guided strategy for engineering antibodies directed against orphan targets or antigens that are difficult to purify was developed and used to humanize the murine MRK-16 monoclonal antibody (mAb). MRK-16 recognizes a conformational epitope of a 170-kDa membrane protein, termed P-glycoprotein (P-gp). Elevated expression of P-gp on tumor cells is associated with resistance to cytotoxic drugs, a major obstacle in chemotherapy. Murine MRK-16 was used to enrich and screen a phage-displayed peptide library to identify reactive mimotopes. One peptide, termed ALR1, was enriched to a greater extent than others and subsequently was expressed as a fusion protein with glutathione S-transferase. ALR1 fusion protein bound MRK-16 specifically and inhibited binding of MRK-16 to cells expressing elevated levels of P-gp. To humanize MRK-16, the murine complementarity determining regions were grafted onto homologous human heavy and light chain variable region frameworks. Framework residues that differed between the murine MRK-16 and the homologous human templates were analyzed and subsequently, five framework positions potentially important for maintaining the specificity and affinity of MRK-16 were identified. A combinatorial library consisting of 32 variants encoding all possible combinations of murine and human residues at the five differing framework positions was expressed in a phage system. In the absence of purified P-gp, ALR1 fusion protein was used as surrogate antigen to screen the antibody library to identify the framework combination that most preserved the binding activity of the mAb. On the basis of the initial screening against the mimotope four antibody variants were selected for further characterization. The binding affinity of these variants for the ALR1 fusion protein correlated with their binding to cells expressing elevated levels of P-gp. Thus, peptide mimotopes which can be identified for virtually any antibody including those that recognize conformational or carbohydrate epitopes, can serve as antigen templates for antibody engineering.

ATP Binding Cassette Transporter, Subfamily B, Mem↗

Human tryptases alpha and beta/II are functionally distinct due, in part, to a single amino acid difference in one of the surface loops that forms the substrate-binding cleft.

Tryptases alpha and beta/II were expressed in insect cells to try to ascertain why human mast cells express these two nearly identical granule proteases. In contrast to that proposed by others, residue -3 in the propeptide did not appear to be essential for the three-dimensional folding, post-translational modification, and/or activation of this family of serine proteases. Both recombinant tryptases were functional and bound the active-site inhibitor diisopropyl fluorophosphate. However, they differed in their ability to cleave varied trypsin-susceptible chromogenic substrates. Structural modeling analyses revealed that tryptase alpha differs from tryptase beta/II in that it possesses an Asp, rather than a Gly, in one of the loops that form its substrate-binding cleft. A site-directed mutagenesis approach was therefore carried out to determine the importance of this residue. Because the D215G derivative of tryptase alpha exhibited potent enzymatic activity against fibrinogen and other tryptase beta/II-susceptible substrates, Asp215 dominantly restricts the substrate specificity of tryptase alpha. These data indicate for the first time that tryptases alpha and beta/II are functionally different human proteases. Moreover, the variation of just a single amino acid in the substrate-binding cleft of a tryptase can have profound consequences in the regulation of its enzymatic activity and/or substrate preference.

Amino Acid Sequence↗

Mechanisms of Surface Precipitation and Dissolution of Barite: A Morphology Approach.

Barite (BaSO4) was synthesized at normal pressure and room temperature by mixing sodium sulfate and barium chloride solutions by gentle flow (0.5 ml/min.). The well-formed rectangular and rhombohedral crystals with smooth faces formed from solutions with low degrees of supersaturation. Most precipitates became single crystals. We employed the scanning electron microscope (SEM) to observe barite dissolution by chemical agent diethylenetrinitrilopentaacetic acid. Molecular modeling was used to assist the interpretation of the surface reaction. A kinetic model for the pit growth (initial dissolution) on the surface was evaluated. With SEM microanalysis of barite and kinetic model of pit growth, important surface phenomena and mechanisms of dissolution can be hypothesized. Surface phenomena and composition are very important in barite dissolution. Experimental results suggest that the initial dissolution starts on the two lowest energy surfaces: (001) and (210), and that the etch pits are elongated in one direction on the (210) surface and formed cavities on the (001) surface. The simulated surface structures by molecular modeling can aid the interpretation the pit formation on those selected surfaces. Copyright 1999 Academic Press.

Journal Article↗

Dynamic norepinephrine alterations in bone marrow: evidence of functional innervation.

Efferent sympathetic nerve activity has been hypothesized to regulate the proliferation and maturation of leukocytes in the bone marrow. Although there is histological evidence for bone marrow innervation and documentation of measurable neurotransmitter, functional activation of these nerves to external stimulation has never been demonstrated. The present study was designed to assess the dynamics of norepinephrine (NE) release in bone marrow in response to well-established protocols known to elevate sympathetic activity. Toward this end, norepinephrine turnover was measured using isotopic and non-isotopic methods in mice in response to cold exposure and bacterial challenge. Cold exposure increased NE turnover rate in bone marrow by 36% from 0.33 to 0.45 ng g(-1) h(-1), while peritoneal Pseudomonas aeruginosa infection increased bone marrow NE turnover rate by 131% from 0.13 to 0.30 ng g(-1) h(-1). These results demonstrate that the adrenergic innervation of the bone marrow is functionally dynamic and is responsive to generalized stress. Furthermore, these results lend credence to the premise that neural mechanisms participate in regulation of lympho- and myelopoietic cellular events.

Animals↗

Preparation and Characterization of Nanoscopic Organic Semiconductor of Oxovanadium Phthalocyanine.

Nanoscopic particles of oxovanadium phthalocyanine (VOPc, phase-II) with different particle sizes are prepared by aggregation of VOPc molecules in the presence of surfactants for the first time. A size effect in the region of nanoscale was observed in the UV-visible spectra. The photoconductivity of the nanoscopic VOPc is much higher than that of bulk VOPc. Copyright 1999 Academic Press.

Journal Article↗

Effect of G40R mutation on the binding of human SRY protein to DNA: a molecular dynamics view.

Molecular dynamics simulation was conducted to investigate the reason why the mutant G40R of hSRY protein has a low affinity for DNA. Compared with the previous dynamics results of the wild-type hSRY-HMG-DNA complex, the results of molecular dynamics simulation on the mutant G40R hSRY-HMG-DNA system demonstrated that the whole structure of DNA (especially the second strand) had a major deviation away from the short arm of the HMG box. Consequently, the DNA and the mutant protein could not specifically recognize each other, that is, very different, and low-occupancy, direct, and water-mediated hydrogen bonds were detected at the protein-DNA interface, no conformational changes occurred at the loop region around Met9 during the simulation, and residue IIe13 did not intercalate between the bases of A5 and A6. These results indicated that the mutant G40R did not form a specific complex with the DNA target, hence led to complete gonadal dysgenesis. From the simulation, we realized that the residue Gly40 played a critical structural role in the hSRY-DNA recognition. It might be a structural supporting point of DNA binding because of the absence of a side chain. The reason for the difficulty of the mutant G40R to form a complex with DNA might be that the long and positively charged side chain of Arg40 by its bulk and positive charge hindered the DNA's access to the active sites of the protein.

DNA-Binding Proteins↗

Protein kinase calpha is an effector of hexamethylene bisacetamide-induced differentiation of Friend erythroleukemia cells.

The program of biochemical and molecular events necessary for commitment to erythroid cell differentiation is particularly well characterized in murine Friend erythroleukemia cell lines. Commitment to hemoglobin synthesis in response to a variety of chemical inducers, including hexamethylene bisacetamide and dimethyl sulfoxide is completed by 24 h and proceeds to terminal differentiation by 96 h. Phorbol 12-myristate 13-acetate, a classical tumor promoter phorbol ester that binds to protein kinase C, blocks differentiation in a reversible manner, suggesting an important role for protein kinase C signaling pathways. The classical protein kinase C isoforms alpha, betaI, and betaII, play distinct roles in the transduction of proliferative and differentiative signals in human, as well as in murine, erythroleukemia cells. Protein kinase Calpha has been implicated in differentiation of human erythroleukemia cells although its translocation to the nucleus has not been observed. Taking advantage of the ability of phorbol 12-myristate 13-acetate to block differentiation in Friend erythroleukemia cells, we determined the localization of the predominant protein kinase C isoforms alpha and betaI during differentiation and in response to their blockade. The ability of phorbol myristate acetate to preferentially diminish protein kinase Calpha-protein localization to the nucleus by 24 h and thereby block differentiation induced by hexamethylene bisacetamide was paralleled by the ability of protein kinase Calpha antisense transfection to block differentiation. In addition, beta-globin transcription, assessed by polymerase chain reaction, was significantly decreased in protein kinase Calpha antisense-transfected cells compared to that seen in vector transfected ones. Taken together, these data suggest an important temporal role for nuclear protein kinase Calpha localization in Friend erythroleukemia cell differentiation.

Acetamides↗

Identification and characterization of immunoreactive calcitonin gene-related peptide from lymphocytes of the rat.

There is accumulating evidence that the immune system can produce neuropeptides. In the light of these facts, we obtained direct evidences to prove that T lymphocytes also synthesize and secrete calcitonin gene-related peptide (CGRP), a neuropeptide localized within primary sensory nerves. By using CGRP specific RIA, CGRP-like immunoreactivity (LI) was found in the extracts of rat lymphocytes from thymus and mesenteric lymph node. The intracellular concentration of lymphocyte-derived CGRP-LI of rat thymus and mesenteric lymph node was 745+/-39 and 447+/-33 fg/10(6) cells, respectively. CGRP-LI in lymphocytes was shown to co-elute with synthetic rat CGRP and sensory neuron-derived CGRP by reverse-phase HPLC. In addition, the CGRP-LI located in the T lymphocytes was also shown by immunocytochemical method examined by electron microscopy. The CGRP mRNA detected by RT-PCR was also present in these lymphocytes and was also identified to be the same one in sensory neurons. These data suggest that CGRP is synthesized and secreted in T lymphocytes of both thymus and lymph node in the rat, and this is identified to be the same one in neuronal tissue. Lymphocyte-derived CGRP may act in an autocrine/paracrine mode and play an important role in certain physiological and pathophysiological conditions.

Animals↗

Hematopoietic progenitor cell abnormalities in Hoxc-8 null mutant mice.

The mammalian Hox genes encode a family of conserved transcription factors that control the establishment of the body plan during embryogenesis. Many Hox genes are also known to be expressed in hematopoietic cells. We found that Hoxc-8, a member of the Hox C cluster, is expressed in the mouse hematopoietic organs, fetal liver and adult bone marrow. To determine the role of Hoxc-8 gene in hematopoiesis, we compared progenitor cell numbers in the fetal liver and adult bone marrow cells. We observed a significant reduction in the number of erythroid burst-forming unit (BFU-E) and in granulocyte/macrophage colony-forming unit (CFU-GM) in the Hoxc-8 null mice, although the peripheral blood cell counts were normal. The hematopoietic cells from the homozygote animals exhibited normal expansion capability in a liquid culture system, suggesting that the decreased number of progenitor cells may be due to a defect extrinsic to the hematopoietic cells, such as in the interaction with the microenvironment.

Amino Acid Sequence↗

Depletion of glutathione by buthionine sulfoxine is cytotoxic for human neuroblastoma cell lines via apoptosis.

Buthionine sulfoximine (BSO) selectively inhibits glutathione (GSH) synthesis and has been used to sensitize tumor cells to alkylating agents, but has minimal single-agent cytotoxicity for most cell types. We determined the cytotoxicity of BSO for 18 (12 MYCN amplified; 6 MYCN nonamplified) human neuroblastoma cell lines using DIMSCAN, a digital image microscopy cytotoxicity assay. D-L(R:S) BSO was highly cytotoxic (>3 logs of cell kill) for most neuroblastoma cell lines, with 17/18 cell lines having IC90 values (range 2. 1->1000 microM) below equivalent steady state plasma levels of L(R:S) BSO reported in adult human trials. Cell lines with genomic amplification of MYCN were more sensitive to BSO than MYCN nonamplified cell lines (P = 0.04). D-L(R:S) BSO (500 microM for 72 h) induced apoptosis as detected by DNA laddering, nuclear morphology, and TUNEL staining of DNA fragments using flow cytometry. Maximal cell killing occurred within 48 h and was antagonized byic value in neuroblastoma.

Antioxidants↗

Measurement of the apparent diffusion coefficient in diffuse renal disease by diffusion-weighted echo-planar MR imaging.

The purpose of this study was to determine the relationship between the apparent diffusion coefficient (ADC) and diffuse renal disease by diffusion-weighted echolanar magnetic resonance (MR) imaging (EPI). Thirty-four patients were examined with diffusion-weighted EPI. The average ADC values were 2.55 x 10(-3) mm2/sec for the cortex and 2.84 x 10(-3) mm2/sec for the medulla in the normal kidneys. The ADC values in both the cortex and medulla in chronic renal failure (CRF) kidneys and in acute renal failure (ARF) kidneys were significantly lower than those of the normal kidneys. In renal artery stenosis kidneys, the ADC values in the cortex were significantly lower than those of the normal and the contralateral kidneys. In the cortex, ADC values were above 1.8 x 10(-3) mm2/sec in all 32 normal kidneys, ranging from 1.6 to 2.0 x 10(-3) mm2/sec in all 8 ARF kidneys, and below 1.5 x 10(-3) mm2/sec in 14 of 15 CRF kidneys. In the medulla, there was considerable overlap in the ADC values of the normal and diseased kidneys. There was a linear correlation between ADC value and sCr level in the cortex (r = 0.75) and a weak linear correlation in the medulla (r = 0.60). Our results show that diffusion-weighted MR imaging may be useful to identify renal dysfunction.

Adolescent↗

Purification of human chorionic gonadotrophin from urine by membrane filtration affinity chromatography with a positively charged membrane.

UNLABELLED: A new method of affinity chromatography, termed membrane filtration affinity chromatography (MFAC), has been developed and applied to purify HCG from urine. By filtrating urine through ZBM (HCG in urine would bind to the antibody on ZBM) and by dissociating the HCG from the antibody on ZBM in purified form, we developed the MFAC and purified HCG from urine of pregnant women by MFAC. The purified HCG showed a single band in polyacrylamide gel electrophoresis. ZBM (1 cm(2)) could harvest 90.3 microg HCG, which showed immunoactivity of 8554 IU/mg. The rate of recovery was 87%. CONCLUSION: MFAC with ZBM is an effective method, which is much easier and cheaper than conventional affinity chromatography for purification of proteins from solution, especially from highly diluted solution.

Chorionic Gonadotropin↗