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The adsorbed conformation of globular proteins at the air/water interface.

External reflection FTIR spectroscopy and surface pressure measurements were used to compare conformational changes in the adsorbed structures of three globular proteins at the air/water interface. Of the three proteins studied, lysozyme, bovine serum albumin and beta-lactoglobulin, lysozyme was unique in its behaviour. Lysozyme adsorption was slow, taking approximately 2.5 h to reach a surface pressure plateau (from a 0.07 mM solution), and led to significant structural change. The FTIR spectra revealed that lysozyme formed a highly networked adsorbed layer of unfolded protein with high antiparallel beta-sheet content and that these changes occurred rapidly (within 10 min). This non-native secondary structure is analogous to that of a 3D heat-set protein gel, suggesting that the adsorbed protein formed a highly networked interfacial layer. Albumin and beta-lactoglobulin adsorbed rapidly (reaching a plateau within 10 min) and with little change to their native secondary structure.

Adsorption↗

Crystal structure of a human CD3-epsilon/delta dimer in complex with a UCHT1 single-chain antibody fragment.

The alpha/beta T cell receptor complex transmits signals from MHC/peptide antigens through a set of constitutively associated signaling molecules, including CD3-epsilon/gamma and CD3-epsilon/delta. We report the crystal structure at 1.9-A resolution of a complex between a human CD3-epsilon/delta ectodomain heterodimer and a single-chain fragment of the UCHT1 antibody. CD3-epsilon/delta and CD3-epsilon/gamma share a conserved interface between the Ig-fold ectodomains, with parallel packing of the two G strands. CD3-delta has a more electronegative surface and a more compact Ig fold than CD3-gamma; thus, the two CD3 heterodimers have distinctly different molecular surfaces. The UCHT1 antibody binds near an acidic region of CD3-epsilon opposite the dimer interface, occluding this region from direct interaction with the TCR. This immunodominant epitope may be a uniquely accessible surface in the TCR/CD3 complex, because there is overlap between the binding site of the UCHT1 and OKT3 antibodies. Determination of the CD3-epsilon/delta structure completes the set of TCR/CD3 globular ectodomains and contributes information about exposed CD3 surfaces.

CD3 Complex↗

Protein component of the ribozyme ribonuclease P alters substrate recognition by directly contacting precursor tRNA.

The protein component of ribonuclease P (RNase P) binds to the RNA subunit, forming a functional ribonucleoprotein complex in vivo and enhancing the affinity of the precursor tRNA (pre-tRNA) substrate. Photocrosslinking experiments with pre-tRNA bound to RNase P reconstituted with the protein component of Bacillus subtilis ribonuclease P (P protein) site specifically modified with a crosslinking reagent indicate that: (i) the central cleft of P protein directly interacts with the single-stranded 5' leader sequence of pre-tRNA, and (ii) the orientation and register of the pre-tRNA leader sequence in the central cleft places the protein component in close proximity to the active site. This unique mode of interaction suggests that the catalytic active site in RNase P occurs near the interface of RNA and protein. In contrast to other ribonucleoprotein complexes where the protein mainly stabilizes the active tertiary fold of the RNA, a critical function of the protein component of RNase P is to alter substrate specificity and enhance catalytic efficiency.

Amino Acid Substitution↗

Mutational analysis of the tetrahydrobiopterin-binding site in inducible nitric-oxide synthase.

Inducible nitric-oxide synthase (iNOS) is a hemeprotein that requires tetrahydrobiopterin (H4B) for activity. The influence of H4B on iNOS structure-function is complex, and its exact role in nitric oxide (NO) synthesis is unknown. Crystal structures of the mouse iNOS oxygenase domain (iNOSox) revealed a unique H4B-binding site with a high degree of aromatic character located in the dimer interface and near the heme. Four conserved residues (Arg-375, Trp-455, Trp-457, and Phe-470) engage in hydrogen bonding or aromatic stacking interactions with the H4B ring. We utilized point mutagenesis to investigate how each residue modulates H4B function. All mutants contained heme ligated to Cys-194 indicating no deleterious effect on general protein structure. Ala mutants were monomers except for W457A and did not form a homodimer with excess H4B and Arg. However, they did form heterodimers when paired with a full-length iNOS subunit, and these were either fully or partially active regarding NO synthesis, indicating that preserving residue identities or aromatic character is not essential for H4B binding or activity. Aromatic substitution at Trp-455 or Trp-457 generated monomers that could dimerize with H4B and Arg. These mutants bound Arg and H4B with near normal affinity, but Arg could not displace heme-bound imidazole, and they had NO synthesis activities lower than wild-type in both homodimeric and heterodimeric settings. Aromatic substitution at Phe-470 had no significant effects. Together, our work shows how hydrogen bonding and aromatic stacking interactions of Arg-375, Trp-457, Trp-455, and Phe-470 influence iNOSox dimeric structure, heme environment, and NO synthesis and thus help modulate the multiple effects of H4B.

Amino Acid Sequence↗

PGDB: a curated and integrated database of genes related to the prostate.

The Prostate Gene Database (PGDB: http://www.ucsf.edu/pgdb) is a curated and integrated database of genes or genomic loci related to the human prostate and prostatic diseases. Currently, PGDB covers genes involved in a number of molecular and genetic events of the prostate including gene amplification, mutation, gross deletion, methylation, polymorphism, linkage and over-expression, as published in the literature. Genes that are specifically expressed in prostate, as evidenced by analysis of data from expressed sequence tags (ESTs) and serial analysis of gene expression (SAGE), are also included. There are a total of 165 unique entries in the database. Users can either browse or query the PGDB through a web interface. For each gene, in addition to basic gene information and rich cross-references to other databases, inclusive and relevant literature references are provided to support the inclusion of the gene in the database. Detailed expression data calculated from the UniGene and SAGEmap databases are also presented.

Databases, Genetic↗

Issues at the clinical-research interface: placebo effect control groups.

At the clinical-research interface, controls are a significant issue in group therapy evaluation. The authors describe and analyze their unique experience with placebo effect control groups conducted prior to brief, group psychotherapy for married couples. Procedures are outlined and participant reactions noted. Solicited ratings and spontaneous comments provide assessment data. The evidence suggests the feasibility of placebo effect control groups in the context studied. Proposed methodological refinements and clinical implications may be useful for future research efforts.

Adult↗

Properties of coralline hydroxyapatite and expanded polytetrafluoroethylene membrane in the immature craniofacial skeleton.

Although extensive research regarding the treatment of calvarial defects has been done in adult models, little is known about the response in the maturing skeleton. The role of coralline hydroxyapatite and expanded polytetrafluoroethylene membrane in augmenting bone growth and repair of calvarial defects in a neonatal model is explored. Utilizing a 3-week-old neonatal swine model, bone growth into 28 calvarial defects was measured. After exposure of the calvaria in seven animals, four defects of 10 mm in diameter were created. In each animal, one defect was treated with a 10-mm disc of porous hydroxyapatite alone (Interpore 500, Interpore International), and a second defect was covered with an expanded polytetrafluoroethylene membrane (Gore-Tex OV-6) secured by four 3-mm microscrews (Luhr Microsystem, Howe-Medica Inc.). The third defect combined an implanted hydroxyapatite disc covered by an expanded polytetrafluoroethylene membrane, whereas the fourth defect served as an untreated control. Histology and histomorphometry were performed on undecalcified specimens harvested at 6 weeks after surgery. In both hydroxyapatite groups, the bone growth into the inorganic matrix provided complete osseous union in all specimens, and the amount of fibrosis was significantly lower (p < 0.02) in comparison with the control. Unexpectedly, there was significant osteoclastic resorption of the hydroxyapatite matrix (35.1 percent decrease) with simultaneous bone deposition and remodeling. The addition of an expanded polytetrafluoroethylene membrane covering the hydroxyapatite implant provided an insignificant advantage in bone growth (27.3 percent versus 28.3 percent, respectively). Finally, the expanded polytetrafluoroethylene membrane alone afforded no qualitative advantage secondary to intrusion of brain and dura into the defect as well as displacement of the membrane inward during appositional growth, leading to incomplete healing of the defect with thinning of the surrounding cranial bone. Unique in this maturing model was morphologic evidence of complete union at the calvaria-hydroxyapatite interface in all specimens as well as active remodeling of the hydroxyapatite matrix. The results of this study suggest that porous hydroxyapatite may be a suitable bone substitute in maturing calvarial bone defects, achieving superior osseous integration and volumetric bone gain while undergoing concurrent resorption and remodeling.

Animals↗

A novel pH-dependent dimerization motif in beta-lactoglobulin from pig (Sus scrofa).

beta-Lactoglobulin (BLG) is a lipocalin and is the major protein in the whey of the milk of cows and other ruminants, but not in all mammalian species. The biological function of BLG is not clear, but a potential role in carrying fatty acids through the digestive tract has been proposed. The capability of BLG to aggregate and form gels is often used to thicken foodstuffs. The structure of the porcine form is sufficiently different from other known BLG structures that SIRAS phases had to be measured in order to solve the crystal structure to 2.4 A resolution. The r.m.s. deviation of C(alpha) atoms is 2.8 A between porcine and bovine BLG. Nevertheless, the typical lipocalin fold is conserved. Compared with bovine BLG, the tilted alpha-helix alters the arrangement of surface residues of the porcine form, completely changing the dimerization behaviour. Through a unique pH-dependent domain-swapping mechanism involving the first ten residues, a novel dimer interface is formed at the N-terminus of porcine BLG. The existence of this novel dimer at low pH is supported by gel-filtration experiments. These results provide a rationale for the difference in physicochemical behaviour between bovine and porcine BLG and point the way towards engineering such dimerization motifs into other members of the lipocalin family.

Amino Acid Motifs↗

A telemetry-instrumentation system for monitoring multiple subcutaneously implanted glucose sensors.

An implantable potentiostat-radiotelemetry system for in vivo sensing of glucose is described. An enzyme electrode sensor measures the oxidation current of hydrogen peroxide formed by the stoichiometric conversion of glucose substrate and oxygen cofactor in an immobilized glucose oxidase layer. The sensor current is converted to a frequency and transmitted at programmable intervals (4, 32, 256 s) to a remote receiver. Low power CMOS circuitry is employed and device operation for up to 1.5 years is predicted using two series connected 250 mAh lithium cells. Crystal controlled RF frequencies uniquely identify each sensor allowing over 10 sensors within the same 10 m radius. A custom interface card allows a PC to program the receiver and handle the transmitted sensor data using software written in Microsoft C and QuickBasic. Software control allows on-the-fly sensor addition or subtraction to the sensor group being monitored. Over 10 sensors can be tracked long-term using the longest transmit interval, or four sensors can be tracked during short-term infusion studies when the transmit interval is reduced to 4 s. The design, construction, operation, and performance of the system hardware and software are described and evaluated.

Animals↗

Episodic tremor and slip on the Cascadia subduction zone: the chatter of silent slip.

We found that repeated slow slip events observed on the deeper interface of the northern Cascadia subduction zone, which were at first thought to be silent, have unique nonearthquake seismic signatures. Tremorlike seismic signals were found to correlate temporally and spatially with slip events identified from crustal motion data spanning the past 6 years. During the period between slips, tremor activity is minor or nonexistent. We call this associated tremor and slip phenomenon episodic tremor and slip (ETS) and propose that ETS activity can be used as a real-time indicator of stress loading of the Cascadia megathrust earthquake zone.

Journal Article↗

Signal pathways and appressorium morphogenesis.

Fungal pathogens have evolved elaborate strategies to gain access to plant tissues. For many pathogens, following attachment of spores to the leaf surface, germ tubes emerge and grow across the surface, often in response to particular environmental cues and to a specific location. At an appropriate site, polar elongation of the germ tube ceases, the tip attaches to the surface and swells to form an appressorium, a uniquely organized infection structure. Following a period of maturation, a hypha then emerges at the plant interface and penetrates into the plant tissues. This chapter discusses recent developments that provide new insight into the molecular mechanism regulating induction and function of appressoria. Topics include attachment to the leaf surface; environmental cues that signal germ-tube growth and appressorium formation; mechanisms for sensing environmental cues; endogenous signaling pathways; and mechanisms of penetration from the appressorium.

Journal Article↗

The evolution of neural circuits controlling feeding behavior in frogs.

Our approach to understanding motor systems is a phylogenetic, 'outside-in' approach, the goal of which is to identify behavioral transitions during phylogenesis and elucidate their neurological basis. In this paper, we review the results of recent behavioral, biomechanical and neurological studies on frog feeding behavior. These studies show that highly protrusible tongues have evolved numerous times independently among frogs, and that the biomechanics and neuromuscular control of feeding behavior have been transformed repeatedly during frog evolution. Many of the independent lineages possess unique biomechanical mechanisms for protracting their tongues and unique neural mechanisms for coordinating feeding behavior. In frogs, there has been considerable evolution at the interface between reticular central pattern generators (CPGs) associated with feeding and sensory feedback circuits that modulate feeding motor output. In particular, the roles of hypoglossal and glossopharyngeal sensory feedback appear to have been relatively plastic in their evolution. Prey-type dependence of hypoglossal sensory feedback in Rana suggests that the interaction between descending visual control and sensory feedback also may be evolutionarily plastic. Comparative studies have found that motor systems sometimes evolve conservatively across morphological and behavioral transitions (i.e., the shoulder in birds) or, alternatively, they may be subject to considerably more evolutionary change than is reflected in morphological characteristics (i.e., feeding in cichlids). We hypothesize that the CPG circuits for feeding behavior in the reticular formation may evolve conservatively because they are highly integrated, multifunctional networks which cannot be optimized for one function without compromising others. In contrast, the interfaces between the CPG, sensory feedback and descending control should be less constrained. When changes in motor patterns occur during evolution, it is likely that sensory feedback or descending control may be involved.

Animals↗

Supporting care homes: the older people's specialist nurse.

This article outlines the health needs of older nursing home residents and identifies the case for the role of an older people's specialist nurse within a multidisciplinary care homes support team (CHST). This model has been successfully introduced in the London boroughs of Lambeth, Southwark and Lewisham as a response to the profile of local health need among care home residents and changes in national policy such as the introduction of NHS-funding nursing care. The structure of the CHST is described, and the older people's specialist nurse is discussed in detail. The emergent role of the older people's specialist nurse as key to managing the interface between the nursing homes and primary care is highlighted as a key benefit of this unique role.

Accidental Falls↗

Maternal/fetal interactions: the role of the MHC class I molecule HLA-G.

The unique pattern of class I major histocompatibility complex (MHC) antigen expression seen at the human maternal/fetal interface is thought to be vital for fetal well-being. The lack of polymorphic class I and II MHC antigens on trophoblasts, the only fetal tissue in direct contact with the mother, is likely to be at least a part of the explanation of fetal evasion of allograft rejection. The recent observation that the HLA-G-encoded class I MHC molecule is present on certain subpopulations of cytotrophoblasts suggests that this nonpolymorphic molecule may have a role in the maternal/fetal immune response. Although no experimental evidence exists to support a particular function for HLA-G, reasoned speculation about the possible roles of this nonpolymorphic class I molecule is possible. Data derived from sequence analysis, analysis of HLA-G expression patterns, analysis of the extraembryonic expression patterns of other genes, and analysis of decidual lymphocyte phenotype and function provide insight into the possible functions of HLA-G at the maternal/fetal interface and are considered here.

Base Sequence↗

Environment and the skin.

The skin is an important interface between man and his environment; it is an important portal of entry for hazardous agents and a vulnerable target tissue as well. It is a uniquely accessible model system for detecting hazards and for studying mechanisms of a wide variety of biologic funcitons. Environmental causes of skin reactions comprise a vast array of physical, chemical and biological agents. To appreciate the role of the skin as an interface with man's environment, it is necessary to understand the multiple adaptive mechanisms, and the defenses of the skin against the environmental stresses. The skin is endowed with a versatile group of defenses against penetration, fluid loss from the body, thermal stress, solar radiation, physical trauma and microbial agents. Patterns of adverse response range in quality and intensity from uncomplicated itching to metastatic neoplasia. Environmental problems comprise a large segment of disabling skin disease. Although critical epidemiologic data is limited, cutaneous illnesses comprise a significant segment of occupational disease. This represents a significant loss in productivity and a major cause of disability. The most serious research needs include the development of surveillance systems for identifying skin hazards and determining frequency of environmental skin disease; the development of new models for studying cutaneous penetration; the elucidation of the mechanisms of nonallergic inflammatory reactions (primary irritation) and of the accommodation phenomenon; the development of more sensitive models for predicting adverse responses to marginal irritants; the utilization of modern skills of immunobiology and immunochemistry to elucidate mechanisms of allergic responses; the launching of epidemiologic studies to determine the long term effects of PCBs and associated compounds such as dioxins; and the expansion of research in the mechanisms of skin cancer in relation to susceptibility, genetic and metabolic considerations, ultraviolet light, and phototoxic agents.

Dermatitis, Contact↗

Binding and transport of metal ions at the dimer interface of the Escherichia coli metal transporter YiiP.

YiiP is a representative member of the cation diffusion facilitator (CDF) family, a class of ubiquitous metal transporters that play an essential role in metal homeostasis. Recently, a pair of Zn2+/Cd2+-selective binding sites has been localized to two highly conserved aspartyl residues (Asp157), each in a 2-fold-symmetry-related transmembrane segment 5 (TM5) of a YiiP homodimer. Here we report the functional and structural interactions between Asp157 and yet another highly conserved Asp49 in the TM2. Calorimetric binding analysis indicated that Asp49 and Asp157 contribute to a common Cd2+ binding site in each subunit. Copper phenanthroline oxidation of YiiP(D49C), YiiP(D157C), and YiiP(D49C/D157C) yielded inter- and intra-subunit cross-links among Cys49 and Cys157, consistent with the spatial proximity of two (Asp49-Asp157) sites at the dimer interface. Hg2+ binding to YiiP(D49C) or YiiP(D49C/D157C) also yielded a Cys49-Hg2+-Cys49 biscysteinate complex across the dimer interface, further establishing the interfacial location of a (Asp49-Asp157)2 bimetal binding center. Two bound Cd2+ ions were found transported cooperatively with a sigmoidal dependence on the Cd2+ concentration (n = 1.4). The binding affinity, transport cooperativity, and rate were modestly reduced by either a D49C or D157C mutation, but greatly diminished when all the bidentate aspartate O-ligands in (Asp49-Asp157)2 were replaced by the monodentate cysteine S-ligands. The functional significance of these findings is discussed based on the unique coordination chemistry of aspartyl residues and a model for the translocation pathway of metal ions at the YiiP dimer interface.

Aspartic Acid↗

Trophinin, tastin, and bystin: a complex mediating unique attachment between trophoblastic and endometrial epithelial cells at their respective apical cell membranes.

Embryo implantation is a complex process consisting of multiple cross-talks between maternal and embryonic cells. Defining the mechanisms underlying implantation at molecular level is challenging task in reproductive biology. In order to identify molecules involved in cellular interactions between trophoblastic and endometrial epithelial cells, we have established two human cell lines, trophoblastic HT-H and endometrial epithelial SNGM. These two cell types exhibit cell adhesion at their respective apical cell membranes. Molecules involved in this unique cell adhesion were identified by expression complementary DNA cloning and were named trophinin, tastin, and bystin. Trophinin is a membrane protein thought to have self-binding activity and thus mediates homophilic cell adhesion. Tastin and bystin are cytoplasmic proteins required for trophinin to exhibit cell adhesion activity. Trophinin is strongly expressed in trophectoderm of monkey blastocysts. In human endometrium, trophinin is expressed for a limited period in the luminal epithelium at the time expected for implantation. In human placenta, trophinin, tastin, and bystin are strongly expressed in trophoblast and endometrium at the uteroplacental interface at an early stage in pregnancy. All these molecules disappear from the human placenta in the second trimester. The unique expression pattern and cell adhesion activity exhibited by trophinin, tastin, and bystin suggest strongly the involvement of these molecules in the initial attachment of blastocyst to uterus.

Animals↗

Synchronized changing of transinterface pressure, bubble radius and surface tension: a unique feature of lung surfactant.

The pulsating bubble surfactometer has been commonly used to measure the minimum surface tension of lung surfactant. The complexity of the original transinterface pressure tracings and its possible physiological meanings remain undefined. In the present study, we compared surface properties between calf lung surfactant extract (CLSE) and Tween 20, a nonionic surfactant, with the pulsating bubble surfactometer. A synchronized change between transinterface pressure (P) and bubble radius (R) was observed when CLSE was tested. Mathematical analysis and computer simulation indicate that this is due to the extremely potent surface tension lowering and adjusting abilities, which allows the surface tension to decrease towards zero at the end of compression and increase towards a high surface tension during re-expansion. In contrast, a time delay between P and R was observed when Tween 20 was assessed. Surface tension adjusting ability was shown only at concentrations below or around the critical micelle concentration (cmc) of Tween 20. Surface tension became unchangeable when concentrations were further increased, suggesting amphipathic molecules were saturated on the interface. The synchronization of transinterface pressure, alveolar radius and surface tension may play an important role in maintaining the pulmonary compliance in vivo. This unique feature, observed at concentrations several orders above the cmc of phospholipids, suggests that the structure of lung surfactant at the air-liquid interface differs from that of Tween 20, a monolayer of free amphipathic molecules.

Chemical Phenomena↗