Biomedical subjects

Qing Deng

Publications and source records attributed to Qing Deng.

8 recordsLinked to original sources

PubMed2007

Multiple WASP-interacting protein recognition motifs are required for a functional interaction with N-WASP.

The WASP-interacting protein (WIP) targets WASP/WAVE proteins through a constitutive interaction with an amino-terminal enabled/VASP homology (EVH1) domain. Parallel investigations had previously identified two distinct N-WASP binding motifs corresponding to WIP residues 451-461 and 461-485, and we determined the structure of a complex between WIP-(461-485) and the N-WASP EVH1 domain (Volkman, B. F., Prehoda, K. E., Scott, J. A., Peterson, F. C., and Lim, W. A. (2002) Cell 111, 565-576). The present results show that, when combined, the WIP-(451-485) sequence wraps further around the EVH1 domain, extending the interface observed previously. Specific contacts with three WIP epitopes corresponded to regions of high sequence conservation in the verprolin family. A central polyproline motif occupied the canonical binding site but in a reversed orientation relative to other EVH1 complexes. This interaction was augmented in the amino- and carboxyl-terminal directions by additional hydrophobic contacts involving WIP residues 454-459 and 475-478, respectively. Disruption of any of the three WIP epitopes reduced N-WASP binding in cells, demonstrating a functional requirement for the entire binding domain, which is significantly longer than the polyproline motifs recognized by other EVH1 domains.

Amino Acid Motifs↗

PubMed2006

Pseudomonas aeruginosa ExoS ADP-ribosyltransferase inhibits ERM phosphorylation.

Pseudomonas aeruginosa causes life-threatening infections in compromised and cystic fibrosis patients. Pathogenesis stems from a number of virulence factors, including four type III translocated cytotoxins: ExoS, ExoT, ExoY and ExoU. ExoS is a bifunctional toxin: the N terminus (amino acids 96-219) encodes a Rho GTPase Activating Protein (GAP) domain. The C terminus (amino acids 234-453) encodes a 14-3-3-dependent ADP-ribosyltransferase domain which transfers ADP-ribose from NAD onto substrates such as the Ras GTPases and vimentin. Ezrin/radixin/moesin (ERM) proteins have recently been identified as high-affinity substrates for ADP-ribosylation by ExoS. Expression of ExoS in HeLa cells led to a loss of phosphorylation of ERM proteins that was dependent upon the expression of ADP-ribosyltransferase activity. MALDI-MS and site-directed mutagenesis studies determined that ExoS ADP-ribosylated moesin at three C-terminal arginines (Arg553, Arg560 and Arg563), which cluster Thr558, the site of phosphorylation by protein kinase C and Rho kinase. ADP-ribosylated-moesin was a poor target for phosphorylation by protein kinase C and Rho kinase, which showed that ADP-ribosylation directly inhibited ERM phosphorylation. Expression of dominant active-moesin inhibited cell rounding elicited by ExoS, indicating that moesin is a physiological target in cultured cells. This is the first demonstration that a bacterial toxin inhibits the phosphorylation of a mammalian protein through ADP-ribosylation. These data explain how the expression of the ADP-ribosylation of ExoS modifies the actin cytoskeleton and indicate that ExoS possesses redundant enzymatic activities to depolymerize the actin cytoskeleton.

ADP Ribose Transferases↗

PubMed2005

Uncoupling Crk signal transduction by Pseudomonas exoenzyme T.

Exoenzyme T (ExoT) is a bifunctional type III cytotoxin of Pseudomonas aeruginosa that possesses both Rho GTPase-activating protein and ADP-ribosyltransferase activities. The ADP-ribosyltransferase activity of ExoT stimulated depolymerization of the actin cytoskeleton independent of Rho GTPase-activating protein function, and ExoT was subsequently shown to ADP-ribosylate Crk (CT10 regulator of kinase)-I and Crk-II. Crk proteins are eukaryotic adaptor proteins comprising SH2 and SH3 domains that are components of the integrin signaling pathway leading to Rac1 and Rap1 functions. Mass spectroscopic analysis identified Arg20 as the site of ADP-ribosylation by ExoT. Arg20 is a conserved residue located within the SH2 domain that is required for interactions with upstream signaling molecules such as paxillin and p130cas. Glutathione S-transferase pull-down and far Western assays showed that ADP-ribosylated Crk-I or Crk-I(R20K) failed to bind p130cas or paxillin. This indicates that ADP-ribosylation inhibited the direct interaction of Crk with these focal adhesion proteins. Overexpression of wild-type Crk-I reduced cell rounding by ExoT, whereas expression of dominant-active Rac1 interfered with the ability of ExoT to round cells. Thus, the ADP-ribosylation of Crk uncouples integrin signaling by direct inhibition of the binding of Crk to focal adhesion proteins.

ADP Ribose Transferases↗

PubMed2003

Aptamer affinity chromatography for rapid assay of adenosine in microdialysis samples collected in vivo.

An anti-adenosine aptamer was evaluated as a stationary phase in packed capillary liquid chromatography. Using an aqueous mobile phase containing 20 mM Mg2+, adenosine was strongly retained on the column. A gradient of increasing Ni2+ (to 18 mM), which is presumed to complex with nitrogen atoms in adenosine involved in binding to the aptamer, eluted adenosine in a narrow zone. Up to 6 microl of 1.2 microM adenosine could be injected onto the 150-microm I.D. x 7 cm long column without loss of adenosine. With UV absorbance detection, the detection limit was 30 nM or 120 fmol (4 microl injected). Samples could be repetitively injected with 4.6% relative standard deviation in peak area. Columns were stable to at least 200 injections. The adenosine assay, which required no sample preparation, was used on microdialysis samples collected from the somatosensory cortex of chloral hydrate anesthetized rats. Total analysis times were short enough that dialysate samples could be injected every 5 min. Basal dialysate concentrations of adenosine stabilized at 87+/-10 nM (n=5) with the probe operated at 0.6 microl/min.

Adenosine↗

PubMed2003

Microscale determination of purines in tissue samples by capillary liquid chromatography with electrochemical detection.

A microscale method for purines involved in intracellular signaling and energy metabolism, including ADP, ATP, cyclic-AMP, NADH and GTP, was developed. The analytes were separated on a fused-silica capillary liquid chromatography column (50 microm inner diameter by 25 cm long) packed with 7 microm reversed-phase particles and detected with a carbon fiber cylinder microelectrode at +1.50 V versus Ag/AgCl reference electrode. With an acetonitrile gradient, the separation was carried out within 15 min. With a 100 nl injection volume, the detection limits varied from 0.9 to 8 fmol depending upon the analyte. The low detection limits make the method suitable for analysis of small tissue samples. As a demonstration of the method, islets of Langerhans were analyzed for their adenosine-related messenger content.

Animals↗

PubMed2003

Isolation, expression pattern of a novel human RAB gene RAB41 and characterization of its intronless homolog RAB41P.

Small GTPases form a big family including Ras, Rho, Rac, Rab, Sar1/Arf subfamilies and Ran homologs, playing important roles in diverse cellular processes. Through data mining, a novel human RAB41 gene was predicted and subsequently isolated from human testis. The open reading frame of RAB41 is 636bp in length. RAB41 was composed of three exons, and it was mapped to chromosome 3q21.3 by comparing with the human genomic data. RAB41 protein contains a RAB domain. Similarity analysis indicated that RAB41 is closely similar to RAB19. The results of PCR amplification indicated that human RAB41 is widely expressed in brain, testis, lung, heart, ovary, colon, kidney, uterus and spleen but not in liver. By genomic searching, an intronless pseudogene homologous to RAB41 at chromosome 16--RAB41P was identified at human chromosome 16q11.2. Flanking the pseudogene RAB41P in human genome, there exist some transposable elements LINEs--L1, whose contribution in the generation of this intronless homolog is discussed.

Amino Acid Sequence↗

PubMed2003

A follow-up study on three caries activity tests.

The purpose of this research was to study the efficacy of three CAT's (Dentocult SM, Dentocult LB and Dentobuff Strip) in revealing caries condition and predicting caries progress, and provide a reference for application by comparing the three tests. Oral condition and results of the three CAT's of 82 children aged 3 to 4 were recorded and followed up. The examination was checked again two years later. The caries incidence, dft and CSI data from the two examinations were analyzed statistically. The results were that each Dentocult SM degree showed significant variances in incidence rate, as did the dft and CSI results in the second examination. The dft and CSI of both examinations exhibited a high degree of statistical significance. The same may be said of the Dentocult LB findings for the two years. No noticeable variances in caries incidence rate, dft and CSI from the Dentobuff Strip test were observed in both years' study, nor was there any statistical significance drawn from the findings, except for those of the second exam. No gender differences were observed in the distribution by degree with the three CATs The conclusion is that Dentocult SM is the best of the three tests for the diagnosis of the presence of caries and prognosis of its progress, Dentocult LB is second best whereas the Dentobuff Strip shows no detection capability. The findings serve as an application reference.

Analysis of Variance↗

PubMed2002

[Study on antiradiation effect of panaxatriol].

OBJECTIVE: To study antiradiation effect of panaxatriol (PT) and to provide an experiment basis for developing relative radiation-resistant skin care products. METHODS: Mice were treated with PT or normal saline (the control group) before X-ray irradiation and were to observe the number of chromosme aberrations killed 3 hours after irradiation. The chromosome samples of bone marrow cells were prepared and chromatid mutation as well as aberrant cells. RESULTS: PT can reduce the frequency of chromatid mutative induced by X-ray. Antiradiation effect was positively correlated with the dose of PT and the duration of drug, and negatively correlated with the dose of X-rays. CONCLUSION: PT has significant antiradiation effect.

Animals↗