Search PubMed⌕ Search

Biomedical subjects

H Tang

Publications and source records attributed to H Tang.

At least 19 recordsLinked to original sources
PubMed2003

Direct determination of the energy required to operate a single molecule switch.

Using a noncontact atomic-force and scanning-tunneling microscope in ultrahigh vacuum, we have measured the switching energy of a single molecule switch based on the rotation of a di-butyl-phenyl leg in a Cu-tetra-3,5 di-tertiary-butyl-phenyl porphyrin. The mechanics and intramolecular conformation of the switched leg is controlled by the tip apex of the noncontact atomic-force microscope. The comparison between experimental and calculated force curves shows that the rotation of the leg requires an energy less than 100 x 10(-21) J, which is 4 orders of magnitude lower than state-of-the-art transistors.

Journal Article↗
PubMed2003

Profiling genes expressed in human fetal cartilage using 13,155 expressed sequence tags.

OBJECTIVE: To analyze the gene expression profile of human fetal cartilage by expressed sequence tags (ESTs). METHODS: A human fetal cartilage (8-12 weeks) cDNA library was constructed using the lambda ZAP Express vector. ESTs were obtained by partial sequencing of cDNA clones. The basic local alignment search tool algorithm was used to compare all generated ESTs to known sequences. RESULTS: A total of 13,155 ESTs were analyzed, of which 8696 ESTs (66.1%) matched known genes, 53 ESTs (0.4%) were putatively novel (with no match) and the rest matched other ESTs, genomic DNA and repetitive sequences. Importantly, we identified 2448 unique known genes through non-redundancy analysis of the known gene matches, which were then functionally categorized. The tissue specificity of this library was reflected by its EST profile of the extracellular matrix (ECM) proteins. Collagens were the major transcripts, representing 68.5% of the ECM proteins. Proteoglycans were the second most abundant, constituting 9.5%. Collagen type II was the most abundant gene of all. Glypican 3, decorin and aggrecan were the major transcripts of proteoglycans. Many genes involved in cartilage development were identified, such as insulin-like growth factor-II, its receptor and binding proteins, connective tissue growth factor and fibroblast growth factors. Proteases and their regulatory factors were also identified, including matrix metalloprotease 2 and tissue inhibitor of metalloproteinase 1. CONCLUSIONS: The EST approach is an effective way of characterizing the genes expressed in cartilage. These data represent the most extensive molecular information on human fetal cartilage to date. The availability of this information will serve as a basis for further research to identify genes that are essential in cartilage development.

Cartilage, Articular↗
PubMed2003

Multiple assembly signals in gamma-aminobutyric acid (type A) receptor subunits combine to drive receptor construction and composition.

Mammalian gamma-aminobutyric acid type A (GABAA) receptors are constructed from a large repertoire of subunits (alpha1-alpha6, beta1-beta3, gamma1-gamma3, delta, epsilon, theta and pi) into a pentameric ion channel. GABA(A) receptor assembly occurs within the endoplasmic reticulum (ER) and involves interactions with chaperone molecules. Only specific subunit combinations can produce functional surface receptors (with a fixed stoichiometry); other subunit combinations are retained within the ER and degraded. Thus, receptor assembly occurs by defined pathways to limit the diversity of GABAA receptors. The key to understanding how receptor diversity is achieved and controlled is the identification of assembly signals capable of distinguishing between other subunit partners. Analysis of an assembly box in alpha1 (residues 57-68) has revealed an absolute requirement for this region in the assembly of alphabeta receptors. Furthermore, a selective requirement for a single amino acid (R66) is observed for the assembly of alpha1beta2, but not alpha1beta1 or alpha1beta3, receptors. In addition, we have characterized an assembly signal in the beta3 subunit that is capable of driving the assembly of beta3, gamma2beta3 and alpha1beta3 receptors. Interestingly, this signal does not appear to utilize the alpha1 assembly box, suggesting the presence of alternative assembly signals within the alpha1 subunit. Although this beta3 signal is sufficient to permit the formation of betagamma receptors it is not necessary, suggesting that alternative assembly signals also exist within the beta3 subunit. These findings support the belief that GABAA receptor assembly occurs via multiple defined pathways that may be determined by subunit availability.

Amino Acid Sequence↗
PubMed2003

Improving the self reporting of tobacco use: results of a factorial experiment.

OBJECTIVES: To examine divergent estimates of smoking prevalence in two random digit dial surveys for the same population. Based upon internal and external reviews of survey procedures, differences in survey introductions (general health versus tobacco specific introduction) and/or differences in the use of filter questions were identified as the most likely explanations. This prompted an experiment designed to investigate these potential sources of measurement error. DESIGN: A randomised 2 x 2 factorial experiment. SETTING: A random digit dial telephone survey from July to September 2000. SUBJECTS: 3996 adult Californian respondents. MAIN OUTCOME MEASURES: A series of smoking prevalence questions in the context of a tobacco or general health survey. RESULTS: Logistic regression analyses suggest that, among females, prior knowledge (from the survey introduction) that a survey is concerned with tobacco use may decrease self reported smoking prevalence (approximately 4% absolute prevalence difference). Differences in the use of filter questions resulted in almost no misclassification of respondents. CONCLUSIONS: The tobacco specific survey introduction is causing some smokers to deny their tobacco use. The data suggest that these smokers tend to be women that smoked occasionally. A desire by the participants to minimise their personal time costs or a growing social disapproval of tobacco use in the USA may be contributing to the creation of previously undetected survey artefacts in the measurement of tobacco related behaviours.

Adolescent↗
PubMed2002

Evidence for the existence of an unfolding intermediate of thyroglobulin during denaturation by guanidine hydrochloride.

The unfolding of bovine thyroglobulin (Tg) in guanidine hydrochloride (GuHCl) solution was studied by following the fluorescence and circular dichroism. With increasing GuHCl concentrations, the emission maximum of the intrinsic fluorescence clearly red-shifted in two stages. At concentrations of GuHCl less than 1.2 M or more than 1.6 M, the red shift showed a cooperative manner. At concentrations of GuHCl between 1.2 and 1.6 M, an unfolding intermediate was observed. It was further characterized by the increased binding of the fluorescence probe 1-anilinonaphthalene-8-sulfonic acid (ANS). No significant changes of the secondary structure were indicated by CD spectra at the concentrations of GuHCl between 1.2 and 1.6 M. The conformation of this state has properties similar to those of a molten globule state which may exist in the folding pathway of the protein. Further changes in fluorescence properties occurred at concentrations of denaturant higher than 1.6 M with a significant red shift of the emission maximum from 340 to 347 nm and a marked decrease in ANS binding. This in vitro study gave a clue to understand the biochemical mechanism for the occurrence of aggregation and molecular chaperone binding during Tg maturation in vivo.

Anilino Naphthalenesulfonates↗
PubMed2001

Crystallization and preliminary X-ray crystallographic studies of a macromolecular antitumour antibiotic, C1027.

C1027 is a macromolecular antitumour antibiotic produced by Streptomyces globispourus C1027 and consists of an apoprotein and a non-protein labile chromophore. Little is known about how the thermally unstable chromophore is stabilized by the apoprotein. The purified C1027 was monodisperse according to dynamic light-scattering measurements and crystallized in two different crystal forms from two different starting conditions using the vapour-diffusion method. Condition I yielded hexagonal prism crystals having space group P3(1)/P3(2) and unit-cell parameters a = b = 66.8, c = 55.4 A. Diffraction data were collected to 2.1 A resolution using an in-house Rigaku rotating Cu anode X-ray generator. Another condition produced rod-like crystals with space group P3(1)21/P3(2)21 and unit-cell parameters a = b = 55.15, c = 55.87 A. A data set to 1.8 A resolution was collected from a rod-like crystal using a MAR CCD detector at the SRS synchrotron source.

Aminoglycosides↗
PubMed2001

Proteins of the cyanobacterial photosystem I.

Cyanobacterial photosystem (PS) I is remarkably similar to its counterpart in the chloroplast of plants and algae. Therefore, it has served as a prototype for the type I reaction centers of photosynthesis. Cyanobacterial PS I contains 11-12 proteins. Some of the cyanobacterial proteins are modified post-translationally. Reverse genetics has been used to generate subunit-deficient cyanobacterial mutants, phenotypes of which have revealed the functions of the missing proteins. The cyanobacterial PS I proteins bind cofactors, provide docking sites for electron transfer proteins, participate in tertiary and quaternary organization of the complex and protect the electron transfer centers. Many of these mutants are now being used in sophisticated structure-function analyses. Yet, the roles of some proteins of the cyanobacterial PS I are unknown. It is necessary to examine functions of these proteins on a global scale of cell physiology, biogenesis and evolution.

Cyanobacteria↗
PubMed2001

Transcription and replication of a natural hepatitis B virus nucleocapsid promoter variant is regulated in vivo by peroxisome proliferators.

A hepatitis B virus (HBV) transgenic mouse containing a naturally occurring mutation in the nucleocapsid promoter (A1764T plus G1766A) that inhibits the retinoid X receptor alpha (RXRalpha) plus peroxisome proliferator-activated receptor alpha (PPARalpha) heterodimer from binding to the proximal nuclear hormone receptor recognition sequence has been generated. Viral transcription and replication occur in the liver and kidney. The nucleocapsid promoter mutation does not prevent peroxisome proliferators from increasing viral transcription and replication in the liver of these variant HBV transgenic mice. This suggests that peroxisome proliferators may enhance viral transcription directly in a PPARalpha-dependent manner through the nuclear hormone receptor recognition site in the enhancer 1 region of the HBV genome. Hepatocyte nuclear factor 4 (HNF4) binding to the proximal nuclear hormone receptor recognition sequence in the nucleocapsid promoter appears to limit RNA synthesis from the precore transcription initiation site. Consequently, the variant HBV transgenic mice transcribe very little precore RNA and secrete extremely low levels of hepatitis B e antigen (HBeAg) compared with the wild-type HBV transgenic mice. This is consistent with the suggestion that viruses expressing HBeAg are preferentially eliminated in infected individuals when they seroconvert from HBeAg positive to anti-HBe antibody-positive status, leaving escape HBV variants that have reduced HBeAg expression.

3T3 Cells↗
PubMed2001

Crystallization and preliminary X-ray analysis of a Trx domain of human thioredoxin-like protein.

The Trx domain of human thioredoxin-like protein has been purified and crystallized using ammonium sulfate as precipitant. The crystal belongs to space group C2, with unit-cell parameters a = 87.5, b = 48.5, c = 29.8 A, beta = 99.59 degrees. It has one molecule per asymmetric unit and diffracts beyond 2.2 A under cryoconditions (100 K) using an in-house Cu rotating-anode X-ray generator.

Crystallization↗
PubMed2001

Crystallization and preliminary X-ray crystallographic analysis of native and selenomethionyl recombinant tabtoxin-resistance protein complexed with acetyl-coenzyme A.

Tabtoxin-resistance protein (TTR), an acetyltransferase from Pseudomonas syringae pv. tabaci, was overexpressed in Eschericha coli M15 and the TTR fusion protein complexed with acetyl-coenzyme A (AcCoA) was purified and crystallized. Diffraction data were collected to 3.0 A resolution in-house and the crystal was found to belong to space group P2(1), with unit-cell parameters a = 47.6, b = 66.6, c = 53.5 A, beta = 104.3 degrees. Furthermore, a selenomethionine (SeMet) TTR fusion protein derivative was overexpressed in the same expression system and its complex with AcCoA was purified in a reductive environment. The SeMet TTR derivative crystallized in two forms: the first was identical to that observed for native crystals and the second belonged to space group C2, with unit-cell parameters a = 101.7, b = 45.6, c = 84.2 A, beta = 105.8 degrees. Data from the P2(1) crystal form were collected in-house to 2.3 A resolution. Subsequently, three different wavelength data sets of the C2 crystal form to 1.55 A resolution were collected at the Advanced Photon Source at Argonne National Laboratory.

Acetyl Coenzyme A↗
PubMed2001

Signal transduction in neuronal migration: roles of GTPase activating proteins and the small GTPase Cdc42 in the Slit-Robo pathway.

The Slit protein guides neuronal and leukocyte migration through the transmembrane receptor Roundabout (Robo). We report here that the intracellular domain of Robo interacts with a novel family of Rho GTPase activating proteins (GAPs). Two of the Slit-Robo GAPs (srGAPs) are expressed in regions responsive to Slit. Slit increased srGAP1-Robo1 interaction and inactivated Cdc42. A dominant negative srGAP1 blocked Slit inactivation of Cdc42 and Slit repulsion of migratory cells from the anterior subventricular zone (SVZa) of the forebrain. A constitutively active Cdc42 blocked the repulsive effect of Slit. These results have demonstrated important roles for GAPs and Cdc42 in neuronal migration. We propose a signal transduction pathway from the extracellular guidance cue to intracellular actin polymerization.

Actins↗
PubMed2001

Introduction to Maxxam all-season passive sampling system and principles of proper use of passive samplers in the field study.

Maxxam all-season passive sampling system (PASS) is introduced in this paper. The PASS can be used to quantitatively and accurately monitor SO2, NO2, O3, and H2S in air in all weather conditions with flexible exposure times from several hours to several months. The air pollution detection limits of PASS are very low. They can be from sub ppb to ppt levels. The principles of proper use of passive samplers in the field study are discussed by using the PASS as an example.

Air Pollutants↗
PubMed2001

An Eulerian path approach to DNA fragment assembly.

For the last 20 years, fragment assembly in DNA sequencing followed the "overlap-layout-consensus" paradigm that is used in all currently available assembly tools. Although this approach proved useful in assembling clones, it faces difficulties in genomic shotgun assembly. We abandon the classical "overlap-layout-consensus" approach in favor of a new euler algorithm that, for the first time, resolves the 20-year-old "repeat problem" in fragment assembly. Our main result is the reduction of the fragment assembly to a variation of the classical Eulerian path problem that allows one to generate accurate solutions of large-scale sequencing problems. euler, in contrast to the celera assembler, does not mask such repeats but uses them instead as a powerful fragment assembly tool.

Algorithms↗
PubMed2001

Recording intramolecular mechanics during the manipulation of a large molecule.

The technique of single atom manipulation by means of the scanning tunneling microscope (STM) applies to the controlled displacement of large molecules. By a combined experimental and theoretical work, we show that in a constant height mode of manipulation the STM current intensity carries detailed information on the internal mechanics of the molecule when guided by the STM tip. Controlling and time following the intramolecular behavior of a large molecule on a surface is the first step towards the design of molecular tunnel-wired nanorobots.

Journal Article↗
PubMed2001

Mapping the functional domains of HAP95, a protein that binds RNA helicase A and activates the constitutive transport element of type D retroviruses.

The complex retroviruses such as human immunodeficiency virus, type 1, employ a virally encoded protein, Rev, to mediate the nuclear export of unspliced and partially spliced mRNA. In contrast, the simian type D retroviruses act through a cis-acting constitutive transport element (CTE) that presumably interacts directly with cellular export proteins. We first reported that RNA helicase A (RHA) is a shuttle protein that binds to functional CTE in vitro and in vivo. Recently, we isolated a novel protein, HAP95, that specifically binds to the nuclear transport domain of RHA and up-regulates CTE-mediated gene expression. Here, using truncation and deletion mutations, we mapped the domains of HAP95 that are important for RHA binding, transactivation of CTE, and nuclear cytoplasmic shuttling. We report evidence for a novel nuclear export signal in HAP95 and showed that the domains involved in RHA binding and nuclear localization are required for CTE activation. Finally, we showed that HAP95 synergizes significantly with RHA on CTE-mediated reporter gene expression and promotes nuclear export of unspliced mRNA in transfected cells. Taken together, these data support the proposal that HAP95 specifically facilitates CTE-mediated gene expression by directly binding to RHA.

Active Transport, Cell Nucleus↗