Biomedical subjects

Weimin Guo

Publications and source records attributed to Weimin Guo.

13 recordsLinked to original sources

PubMed2006

Proteasome-dependent regulation of signal transduction in retinal pigment epithelial cells.

As in many other types of cells, retinal pigment epithelial (RPE) cells have an active ubiquitin-proteasome pathway (UPP). However, the function of the UPP in RPE remains to be elucidated. The objective of this study is to determine the role of the UPP in controlling the levels and activities of transcription factors hypoxia-inducible factor (HIF) and NF-kappaB. We inhibited the UPP with proteasome-specific inhibitors and determined the activation of HIF and NF-kappaB as well as the expression and secretion of pro-angiogenic factors. HIF-1alpha was not detectable in ARPE-19 cells under normal culture conditions. However, when proteasome activity was inhibited, HIF-1alpha accumulated in RPE in a time-dependent manner. Consistent with accumulation of HIF-1alpha in the cells, levels of mRNA for vascular endothelial growth factor (VEGF) and angiopoietin-2 (Ang-2) in RPE were up to 7-fold higher upon inhibition of the proteasome. Proteasome inhibition was also associated with a 2-fold increase in levels of mRNA for angiopoietin-1 (Ang-1). ARPE-19 cells secrete significant levels of VEGF under normal culture conditions. Inhibition of proteasome activity increased the secretion of VEGF by 2-fold. In contrast to the increase in HIF activity, NF-kappaB activation was reduced by proteasome inhibition. In addition, the expression and secretion of monocyte chemoattractant protein-1 (MCP-1) by RPE were substantially attenuated by the inhibition of proteasome activity. These data demonstrate that the UPP plays an important role in modulating the activities of HIF and NF-kappaB in the RPE. Consequences of an impairment of the UPP include accumulation of HIF-1alpha and diminished NF-kappaB activation, which lead to enhanced expression and secretion of pro-angiogenic factors and attenuated expression of MCP-1. Taken together, these data predict that the impairment of the UPP could lead to the development of AMD-related phenotypes.

Angiogenesis Inducing Agents↗

PubMed2006

Green tea catechin, epigallocatechin-3-gallate, inhibits vascular endothelial growth factor angiogenic signaling by disrupting the formation of a receptor complex.

A potential mechanism by which green tea may prevent cancer development is through the inhibition of angiogenesis. We have shown previously that the green tea catechin, epigallocatechin gallate (EGCG), inhibits endothelial cell tube formation through the inhibition of vascular endothelial growth factor (VEGF)-induced Akt activation and vascular endothelial (VE)-cadherin phosphorylation. Furthermore, EGCG can suppress oxidant-induced production of the proangiogenic cytokine interleukin (IL)-8. To further elucidate the antiangiogenic mechanisms of EGCG, we investigated its regulation of other molecular processes in VEGF-induced signaling in human umbilical vein endothelial cells (HUVECs). We show that EGCG at physiological doses (0.5-10 microM) markedly inhibits the formation of a vascular endothelial growth factor receptor 2 complex formed upon the binding of its ligand VEGF. This disruption results in a significant and dose-dependent decrease in PI3-kinase activity. Electrophoretic mobility shift assay revealed that EGCG decreased the PI3 kinase-dependent activation and DNA-binding ability of NF-kappaB, likely acting through decreasing phosphorylation and degradation of IkappaB. VEGF-induced IL-8 production at the mRNA (real time RT-PCR) and protein levels (ELISA) are also suppressed with EGCG. These results suggest a novel mechanism for green tea's anticancer effects where EGCG can abrogate VEGF signaling by interfering with the formation of a receptor complex, resulting in attenuated mitogenic and angiogenic signaling.

Catechin↗

PubMed2006

Quantitative structure-activity relationship studies on HEPTs by supervised stochastic resonance.

Quantitative structure-activity relationship studies (QSAR) on HEPTs were performed by using a new approach--supervised stochastic resonance (SSR) in this paper. Errors in physicochemical properties have great effects on variable selection and the predictive capability of QSAR models but errors-in-variables were seldom discussed in QSAR. In this paper, based on the theory of stochastic resonance (SR), SSR was proposed and employed to the problem. In SSR, errors and abundant variables were regarded as noise and the relevant descriptors as signals. In the nonlinear systems involved in the SR, the signal and the noise interact harmonically and the signal was consequently enhanced. Therefore, the correlation between the relevant variables and a specified activity of a series molecule was improved by SSR. It is demonstrated that the obtained QSAR models for HEPT analogues by SSR were comparable to those by published methods in their stability and predictivity. SSR is an efficient and promising approach to QSAR studies.

Molecular Structure↗

PubMed2006

The triage of damaged proteins: degradation by the ubiquitin-proteasome pathway or repair by molecular chaperones.

Accumulation of damaged proteins is causally related to many age-related diseases. The ubiquitin-proteasome pathway (UPP) plays a role in selective degradation of damaged proteins, whereas molecular chaperones, such as heat shock proteins, are involved in refolding denatured proteins. This work demonstrates for the first time that the UPP and molecular chaperones work in a competitive manner and that the fates of denatured proteins are determined by the relative activities of the UPP and molecular chaperones. Enhanced UPP activity suppresses the refolding of denatured proteins whereas elevated chaperone activity inhibits the degradation of denatured proteins. CHIP, a co-chaperone with E3 activity, plays a pivotal role in determining the fates of the damaged proteins. The delicate balance between UPP-mediated degradation and refolding of denatured proteins is governed by relative levels of CHIP and other molecular chaperones. Isopeptidases, the enzymes that reverse the actions of CHIP, also play an important role in determining the fate of denatured proteins.

Animals↗

PubMed2006

Novel distance-based atom-type topological indices DAI for QSPR/QSAR studies of alcohols.

In this work, we propose a distance-based atom-type topological index (DAI) for quantitative structure-property/activity relationship (QSPR/QSAR) studies. The newly constructed index, which codes the structural environment of each atom type in a molecule, can be calculated simply. These atom-type topological indices, along with our recently proposed Lu index, were used to construct QSPR/QSAR models for several representative physical properties and biological activities of several data sets of alcohols with a range of non-hydrogen atoms by using multiple linear regression (MLR) analysis. The efficiency of these indices is verified by high quality QSPR models. The results indicate that the combined use of Lu and DAI indices promises to be a useful method for QSPR/QSAR analysis of complex compounds.

Alcohols↗

PubMed2006

Ubiquitin-proteasome pathway function is required for lens cell proliferation and differentiation.

PURPOSE: The ubiquitin proteasome pathway is involved in the regulation of many cellular processes, such as cell cycle control, signal transduction, transcription, and removal of obsolete proteins. The objective of this work was to investigate roles for this proteolytic pathway in controlling the differentiation of lens epithelial cells into lens fibers. METHODS: bFGF-induced cell proliferation was monitored in rat lens epithelial explants by bromodeoxyuridine (BrdU) incorporation. Indicators of lens differentiation included expression of crystallins, lens major intrinsic protein 26 (MIP26), CP49, and filensin and morphologic changes such as cell multilayering and elongation or loss of nuclei. Clasto-lactacystin-beta-lactone, the proteasome-specific inhibitor, was used to study the role of the proteasome in controlling the proliferation and differentiation processes. RESULTS: Explants treated with bFGF initially underwent enhanced proliferation, as indicated by BrdU incorporation and multilayering of the epithelial cells. By 4 days of bFGF treatment, most cells withdrew from the cell cycle, as indicated by diminished BrdU incorporation. After 7 days of treatment with bFGF, lens epithelial explants displayed characteristics of lens fibers, including higher ratios of crystallins to other cytoplasmic proteins and expression of large quantities of MIP26, CP49, and filensin. Adding the proteasome inhibitor to the medium simultaneously with bFGF (day 0) or at day 4 prohibited or delayed bFGF-induced cell proliferation and differentiation. This was indicated by reduced BrdU incorporation and decreased expression of beta- and gamma-crystallins, MIP26, CP49, and filensin. Proteasome inhibition also significantly decreased the number of layers and the sizes of differentiating fibers. CONCLUSIONS: These data show that proteasome activity is required not only for lens cell proliferation but also required for the transition from the epithelial phenotype to the fiber phenotype.

Animals↗

PubMed2006

Ubiquitin-conjugating enzyme 3 delays human lens epithelial cells in metaphase.

PURPOSE: Ubc3/Cdc34 is a ubiquitin-conjugating enzyme (Ubc) with well established functions in the G1-to-S-phase transition. Expecting to find similar effects in human lens epithelial cells (HLECs), the authors explored roles for this ubiquitin-conjugating enzyme in regulation of the HLEC cycle. METHODS: Catalytically incompetent Ubc3 (C88S, L97S), wild-type (wt)Ubc3, and mutant (mt)Ubc2 (C93A) were expressed in HLECs, by using an adenoviral vector, and cell cycle progression was assessed. RESULTS: Expression of mt- and wtUbc3, but not empty virus or mtUbc2, delayed the cell cycle in metaphase, rather than the expected G1 phase. Expression of both Ubc3s also stabilized M-phase regulators, cyclin A, cyclin B, and securin. Thus, it appeared that the Ubc3 enzymes were playing roles different from canonical proteolytic functions in targeting G1/S regulators for degradation. We also directly investigated the effect of inhibiting the proteasome on the cell cycle of HLECs. When the proteasome inhibitor was added to S-phase cells, the M-phase regulators were stabilized, and the cells were arrested in the G2/M phase. In contrast, if the proteasome inhibitor was added before the cells entered the S phase, stabilization of the G1 kinase inhibitors p21WAF and p27KIP was observed and the cells were arrested in the G1 phase. CONCLUSIONS: The ubiquitin-proteasome pathway is involved in regulation of transitions between all phases of the HLEC cycle. However, in contrast with previously described roles for Ubc3 in governing G1/S transitions, expression of Ubc3 delays the HLEC cycle in metaphase. The data suggest novel roles for Ubc3 that do not involve the transfer of ubiquitin in the M phase in the HLEC cell cycle.

Adenoviridae↗

PubMed2005

QSPR study on soil sorption coefficient for persistent organic pollutants.

Quantitative structure-property relationship (QSPR) models of soil sorption coefficients for 32 persistent organic pollutants were constructed using our recently introduced Lu index and novel distance-based atom-type DAI topological indices. Using multiple linear regression technique, a 6-variable model was obtained with the correlation coefficient of estimations (R) being 0.95, and the standard error of estimations (s) being 0.23, and the correlation coefficient (R(cv)) and the standard error (s(cv)) in the leave-4-out cross-validation procedure are 0.90 and 0.31, respectively. The results in this study indicate that soil sorption coefficients of POPs are dominated by molecular size while some DAI indices have smaller influence.

Adsorption↗

PubMed2005

Chemical structure of flavonols in relation to modulation of angiogenesis and immune-endothelial cell adhesion.

The antioxidant activity of flavonoids has been suggested to contribute to several health benefits associated with the consumption of fruits and vegetables. Four flavonols - myricetin (M), quercetin (Q), kaempferol (K) and galangin (G), all with different numbers of hydroxyl moieties (-OH) - were examined for their antioxidant activity and cytotoxicity on human umbilical vein endothelial cells (HUVECs) and for their potential antiangiogenic and cell adhesion effects. The relative antioxidant capacity of these flavonols in cell culture medium (cell-free system) and their intracellular antioxidant activity were M = Q > K = G, which correlated respectively with the presence of 3, 2, 1 and 0 moieties of -OH on their B-ring. The higher the numbers of -OH moieties on the B-ring the less toxic the flavonol was to HUVEC, and the LD50 was determined as: M (100 microM) > Q (50 microM) > K (20 microM) > G (10 microM). These flavonols at approximately 0.5 LD50 doses suppressed the vascular endothelial growth factor (VEGF)-stimulated HUVEC tubular structure formation by: M (47%) > Q (37%) > K (15%) > G (14%), which was not linearly associated with their numbers of -OH moieties. However, the magnitude of flavonols' suppression of activated U937 monocytic cells adhesion to HUVEC was associated with the number of -OH moieties on the B-ring. This was prominent when U937 cells were pretreated with these flavonols. In contrast, the numbers of -OH moiety had no apparent influence on the adhesion or expression of adhesion molecules when activated HUVECs were pretreated with these flavonols. The presence of different numbers of -OH moieties on the B-ring of the flavonols may contribute to their antioxidant activity as well as their toxicity and may play an important role in their potency for biological action such as angiogenesis and immune-endothelial cell adhesion, which, respectively, are important processes in the development of cancer and atherosclerosis.

Antioxidants↗

PubMed2005

Lys6-modified ubiquitin inhibits ubiquitin-dependent protein degradation.

Ubiquitin plays essential roles in various cellular processes; therefore, it is of keen interest to study the structure-function relationship of ubiquitin itself. We investigated the modification of Lys(6) of ubiquitin and its physiological consequences. Mass spectrometry-based peptide mapping and N-terminal sequencing demonstrated that, of the 7 Lys residues in ubiquitin, Lys(6) was the most readily labeled with sulfosuccinimidobiotin. Lys(6)-biotinylated ubiquitin was incorporated into high molecular mass ubiquitin conjugates as efficiently as unmodified ubiquitin. However, Lys(6)-biotinylated ubiquitin inhibited ubiquitin-dependent proteolysis, as conjugates formed with Lys(6)-biotinylated ubiquitin were resistant to proteasomal degradation. Ubiquitins with a mutation of Lys(6) had similar phenotypes as Lys(6)-biotinylated ubiquitin. Lys(6) mutant ubiquitins (K6A, K6R, and K6W) also inhibited ATP-dependent proteolysis and caused accumulation of ubiquitin conjugates. Conjugates formed with K6W mutant ubiquitin were also resistant to proteasomal degradation. The dominant-negative effect of Lys(6)-modified ubiquitin was further demonstrated in intact cells. Overexpression of K6W mutant ubiquitin resulted in accumulation of intracellular ubiquitin conjugates, stabilization of typical substrates for ubiquitin-dependent proteolysis, and enhanced susceptibility to oxidative stress. Taken together, these results show that Lys(6)-modified ubiquitin is a potent and specific inhibitor of ubiquitin-mediated protein degradation.

Adenosine Triphosphate↗

PubMed2004

Regulation of the ubiquitin proteasome pathway in human lens epithelial cells during the cell cycle.

Most proliferating cells follow a series of orderly transitions from one phase to another. These transitions are usually controlled by timed degradation of cell cycle regulators by the ubiquitin-proteasome pathway (UPP). There are no published reports regarding the timing of phases of the human lens cell cycle or regarding cell cycle-related changes in UPP components. Objectives of this study were to characterize the timing of the phases of the human lens epithelial cell cycle and to explore potential functions of critical components of the UPP in controlling lens cell cycle. Human lens epithelial cells were synchronized at G0/G1 phase by contact inhibition. Cell cycle progression upon subculturing was monitored by FACS analysis. It took approximately 40 hr for HLEC to complete one cell cycle, approximately 20 hr for G1 phase, approximately 8-10 hr for S phase and approximately 10 hr for the combination of G2 and M phases. Proteasome-dependent degradation of p21WAF and p27Kip, the dominant Cdk inhibitors, was associated with the G1/S phase transition in these cells. Proteasome inhibition experiments indicate that proteolysis is the predominant process which is responsible for the variations in these regulators during the cell cycle. Levels of specific ubiquitin conjugating enzymes, Ubc7 and Ubc10, increased 6 and 2-fold at the G2/M phase and S/G2/M phases, respectively. Levels of these E2s decreased precipitously upon completion of the M phase. In contrast, levels of ubiquitin activating enzyme (E1) and Ubc3 remained constant during the cell cycle. Cul1, a component of the SCF (an E3), remained relatively constant during cell cycle. The up-regulation of Ubc7 and Ubc10 during the G2/M and S/G2/M phases suggests that these enzymes may be involved in controlling the cell cycle progression at this phase. Taken together, the data indicate that expression of key components of the UPP in the human lens epithelial cells is regulated in a cell cycle-dependent manner. Some of the variations in levels of ubiquitin conjugating enzymes are suggestive of previously undescribed functions.

Cell Cycle↗

PubMed2004

Differential regulation of components of the ubiquitin-proteasome pathway during lens cell differentiation.

PURPOSE: To investigate the role for the ubiquitin-proteasome pathway in controlling lens cell proliferation and differentiation and the regulation of the ubiquitin conjugation machinery during the differentiation process. METHODS: bFGF-induced lens cell proliferation and differentiation was monitored in rat lens epithelial explants by bromodeoxyuridine (BrdU) incorporation and expression of crystallins and other differentiation markers. Levels of typical substrates for the ubiquitin-proteasome pathway, p21(WAF) and p27(Kip), were monitored during the differentiation process, as were levels and activities of the enzymes involved in ubiquitin conjugation. RESULTS: Explants treated with bFGF initially underwent enhanced proliferation as indicated by BrdU incorporation. Then they withdrew from the cell cycle as indicated by diminished BrdU incorporation and accumulation of p21(WAF) and p27(Kip). bFGF-induced cell proliferation was prohibited or delayed by proteasome inhibitors. Lens epithelial explants treated with bFGF for 7 days displayed characteristics of lens fibers, including expression of large quantities of crystallins. Whereas levels of E1 remained constant during the differentiation process, the levels of ubiquitin-conjugating enzyme (Ubc)-1 increased approximately twofold, and the thiol ester form of Ubc1 increased approximately threefold on 7 days of bFGF treatment. Levels of Ubc2 increased moderately on bFGF treatment, and most of the Ubc2 was found in the thiol ester form. Although levels of total Ubc3 and -7 remained unchanged, the proportions of Ubc3 and -7 in the thiol ester form were significantly higher in the bFGF-treated explants. Levels of Ubc4/5 and -9 also increased significantly on treatment with bFGF, and more than 90% of Ubc9 was found in the thiol ester form in the bFGF-treated explants. In contrast, levels of Cul1, the backbone of the SCF type of E3s, decreased 50% to 70% in bFGF-treated explants. CONCLUSIONS: The data show that proteolysis through the ubiquitin-proteasome pathway is required for bFGF-induced lens cell proliferation and differentiation. Various components of the ubiquitin-proteasome pathway are differentially regulated during lens cell differentiation. The downregulation of Cul1 appears to contribute to the accumulation of p21(WAF) and p27(Kip), which play an important role in establishing a differentiated phenotype.

Animals↗

PubMed2002

Prediction and application in QSPR of aqueous solubility of sulfur-containing aromatic esters using GA-based MLR with quantum descriptors.

Quantitative structure-property relationships (QSPR) were developed using a genetic algorithm (GA)-based variable-selection approach with quantum chemical descriptors derived from AM1-based calculations (MOPAC7.0). With the QSPR models, the aqueous solubility of 71 aromatic sulfur-containing carboxylates, including phenylthio, and phenylsulfonyl carboxylates were efficiently estimated and predicted. Using GA-based multivariate linear regression (MLR) with cross-validation procedure, the most important descriptors were selected from a pool of 28 quantum chemical semi-empirical descriptors, including steric and electronic types, to build QSPR models. The molecular descriptors included molecular surface (SA), charges on carboxyl group (Q(oc)), the magnitude of the difference between E(HOMO) of the solute and ELUMO of water, divided by 100 (E(B)), which were main factors affecting the aqueous solubility of the compounds of interest. The resulted coefficients R and R2 of 0.9571 and 0.9161 and the prediction residual error sum of squares (PRESS) of 13.1768, revealed that it was accurate and reliable for the model to predict the aqueous solubility of the investigated organic compounds. If two outliers were omitted from the dataset, the resulted coefficients R = 0.9619, R2 = 0.9253, and PRESS = 10.3875 were significantly improved. Compared with stepwise regression analysis, the results obtained in this work were better and more reasonable. The best QSPR model were obtained by GA-based MLR. Reasonable mechanisms for aqueous solubility of the sulfur-containing carboxylates were investigated and interpreted.

Algorithms↗