Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “modulators”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 1,585 records · Page 88Linked to original sources
PubMed2002

Dose reduction in multi-slice CT of the heart by use of ECG-controlled tube current modulation ("ECG pulsing"): phantom measurements.

PURPOSE: To evaluate the effect of ECG-controlled tube current modulation on radiation exposure in retrospectively-ECG-gated multislice CT (MSCT) of the heart. MATERIAL AND METHODS: Three different cardiac MSCT protocols with different slice collimation (4 x 1, and 4 x 2.5mm), and a pitch-factor of 1.5 and 1.8 were investigated at a multi-slice CT scanner Somatom Volume Zoom, Siemens. An anthropomorphic Alderson-Rando phantom was equipped with LiF-thermoluminescence dosimeters at several organ sites, and effective doses were calculated using ICRP-weighting factors. Scan protocols were performed with ECG-controlled tube current modulation ("ECG pulsing") at two different heart rates (60 and 80 bpm). These data were compared to previous data from MSCT of the heart without use of "ECG pulsing". RESULTS: Radiation exposure with (60 bpm) and without tube current modulation using a 2.5 mm collimation was 1.8 mSv and 2.9 mSv for females, and 1.5 mSv and 2.4 mSv for males, respectively. For protocols using a 1 mm collimation with a pitch-factor of 1.5 (1.8), radiation exposure with and without tube current modulation was 5.6 (6.3) mSv and 9.5 (11.2) mSv for females, and 4.6 (5.2) mSv and 7.7 (9.2) mSv for males, respectively. At higher heart rates (80 bpm) radiation exposure is increased from 1.5 - 1.8 mSv to 1.8 - 2.1 mSv, using the 2.5 mm collimation, and from 4.6 - 5.6 mSv to 5.9 - 7.2 mSv, for protocols using 1 mm collimation. CONCLUSIONS: The ECG-controlled tube current modulation allows a dose reduction of 37 % to 44 % when retrospectively ECG-gated MSCT of the heart is performed. The tube current - as a function over time - and therefore the radiation exposure is dependent on the heart rate.

Coronary Angiography↗
PubMed2002

Minimally invasive hepatectomy: modulation of systemic reactions to operation or laparoscopic approach?

BACKGROUND: A new concept of surgical stress has been proposed that consists of both aggressiveness of operation and systemic reactions to an operation. METHODS AND RESULTS: We have investigated a possible modulation of such systemic reactions to operation and have demonstrated the following 3 points: (1) coagulation and fibrinolytic systems are independently activated during hepatectomy and such activation can be modulated by protease inhibitors such as nafamostat mesilate and antithrombin III; (2) elevated thromboxane A2 during hepatectomy is characterized in the prostanoid system, the elevation of thromboxane A2 is inhibited by thromboxane A2 synthetase inhibitor, and postoperative liver injury is reduced; (3) cytokine response induced by hepatectomy is modulated by preoperative administration of methylprednisolone, leading to possible prevention of bacterial translocation. Therefore, modulating systemic reactions to hepatectomy may be important for successful minimally invasive hepatectomy. Another important option for minimally invasive hepatectomy is the use of operative procedures such as laparoscope or thoracoscope. We have investigated the usefulness of a laparoscopic hepatectomy from the standpoints of early and long-term outcome after hepatectomy. Laparoscopic hepatectomy, which is a difficult and dangerous procedure, can be a feasible option and can result in better short-term outcome and a similar long-term outcome after hepatectomy when compared with conventional open hepatectomy. Therefore, the laparoscopic approach is also a viable option for minimally invasive hepatectomy. CONCLUSIONS: Modulation of systemic reactions to the operation itself and laparoscopic hepatectomy may be new strategies for performing minimally invasive hepatectomy.

Blood Coagulation↗
PubMed2004

Masking and scrambling in the auditory thalamus of awake rats by Gaussian and modulated noises.

This paper provides a look at how modulated broad-band noises modulate the thalamic response evoked by brief probe sounds in the awake animal. We demonstrate that noise not only attenuates the response to probe sounds (masking) but also changes the temporal response pattern (scrambling). Two brief probe sounds, a Gaussian noise burst and a brief sinusoidal tone, were presented in silence and in three ongoing noises. The three noises were targeted at activating the auditory system in qualitatively distinct ways. Dynamic ripple noise, containing many random tone-like elements, is targeted at those parts of the auditory system that respond well to tones. International Collegium of Rehabilitative Audiology noise, comprised of the sum of several simultaneous streams of Schroeder-phase speech, is targeted at those parts of the auditory system that respond well to modulated sounds but lack a well defined response to tones. Gaussian noise is targeted at those parts of the auditory system that respond to acoustic energy regardless of modulation. All noises both attenuated and decreased the precise temporal repeatability of the onset response to probe sounds. In addition, the modulated noises induced context-specific changes in the temporal pattern of the response to probe sounds. Scrambling of the temporal response pattern may be a direct neural correlate of the unfortunate experience of being able to hear, but not understand, speech sounds in noisy environments.

Acoustic Stimulation↗
PubMed2003

Dietary zinc modulates gene expression in murine thymus: results from a comprehensive differential display screening.

Differential mRNA display was used to comprehensively screen the murine thymic transcriptome for genes modulated in vivo by dietary zinc. A moderate feeding protocol rendered young adult, outbred mice zinc-deficient and zinc-supplemented without alterations in feeding behavior or growth. However, these levels of deficiency and supplementation altered specific mRNA abundances in a manner detectable by differential display. In total, 240 primer-pair combinations were used to generate >48,000 interpretable cDNA bands derived from thymic total RNA, of which only 265 or 0.55% were identified as zinc-modulated under these moderate dietary conditions. The most strongly zinc-modulated cDNAs identified by display were reamplified and sequenced. No cDNAs encoding zinc-metalloenzymes or zinc-finger transcription factors were identified as zinc-modulated in this global screening. Those zinc-regulated genes independently confirmed by quantitative PCR included: heat shock proteins 40 and 60; heat shock cognate 70; histocompatibility 2, class II antigen A, alpha; and the T cell cytokine receptor. In addition, a variety of transcription- and translation-related factors (such as ribosomal proteins L3, L5, and L28; nuclear matrix protein 84; matrin cyclophilin; the H3 histone family 3A protein; beta(2) microglobulin; and a cleavage and polyadenylation factor) were identified as zinc-modulated. These profiling data show that differential expression of genes in the thymus in response to the dietary zinc supply precedes many of the phenotypic effects on thymic function associated with severe zinc restriction or supplementation. Several genes involved in T cell development were identified as regulated by zinc and will be targets to evaluate the effects of zinc on immune function.

Animals↗
PubMed2006

Attentional modulation of sensorimotor processes in the absence of perceptual awareness.

Attention modulates visual perception and is generally considered inextricably linked with conscious awareness: we become aware of stimuli as we attend to them, and we attend to stimuli as we become aware of them. Recent evidence suggests that attention can also modulate the effects of stimuli that remain invisible, and a natural explanation is that attention enhances weak perceptual representations, bringing them closer to conscious threshold even if they do not reach that threshold. However, there is also the possibility that attention may modulate neural processes that are entirely separate from those supporting conscious perception: sensorimotor mechanisms that do not create awareness however much they are enhanced. Here we provide evidence in support of this second hypothesis by showing that attentional cueing can modulate the behavioral response to invisible stimuli in a way that is distinct from enhancing their visibility. We used a masked-prime paradigm that produces a negative or positive compatibility effect depending on the perceptual strength (duration or brightness) of the prime. We found that attention enhanced the effect of both visible and invisible primes and also increased the likelihood of detecting the prime (i.e., boosted perceptual strength). Crucially, the pattern of attentional influence on priming could not be explained by attentional modulation of the prime's perceptual strength but was predicted by a direct attentional influence on the nonconscious priming process itself. Therefore, in addition to regulating what we perceive, attention seems to influence our behavior through sensorimotor processes that are not involved in conscious awareness.

Attention↗
PubMed2006

Identification of inflammatory gene modules based on variations of human endothelial cell responses to oxidized lipids.

Oxidized phospholipids are thought to promote atherogenesis by stimulating endothelial cells (ECs) to produce inflammatory cytokines, such as IL-8. In studies with mouse models, we previously demonstrated that genetic variation in inflammatory responses of endothelial cells to oxidized lipids contributes importantly to atherosclerosis susceptibility. We now show that similar variations occur in cultured aortic ECs derived from multiple heart transplant donors. These variations were stably maintained between passages and, thus, reflect either genetic or epigenetic regulatory differences. Expression array analysis of aortic EC cultures derived from 12 individuals revealed that >1,000 genes were regulated by oxidized phospholipids. We have used the observed variations in the sampled population to construct a gene coexpression network comprised of 15 modules of highly connected genes. We show that several identified modules are significantly enriched in genes for known pathways and confirm a module enriched for unfolded protein response (UPR) genes using siRNA and the UPR inducer tunicamycin. On the basis of the constructed network, we predicted that a gene of unknown function (MGC4504) present in the UPR module is a target for UPR transcriptional activator ATF4. Our data also indicate that IL-8 is present in the UPR module and is regulated, in part, by the UPR. We validate these by using siRNA. In conclusion, we show that interindividual variability can be used to group genes into pathways and predict gene-gene regulatory relationships, thus identifying targets potentially involved in susceptibility to common diseases such as atherosclerosis.

Activating Transcription Factor 4↗
PubMed2002

In vivo modulation of nonphotochemical exciton quenching (NPQ) by regulation of the chloroplast ATP synthase.

Nonphotochemical quenching (NPQ) of excitation energy, which protects higher plant photosynthetic machinery from photodamage, is triggered by acidification of the thylakoid lumen as a result of light-induced proton pumping, which also drives the synthesis of ATP. It is clear that the sensitivity of NPQ is modulated in response to changing physiological conditions, but the mechanism for this modulation has remained unclear. Evidence is presented that, in intact tobacco or Arabidopsis leaves, NPQ modulation in response to changing CO(2) levels occurs predominantly by alterations in the conductivity of the CF(O)-CF(1) ATP synthase to protons (g(H)(+)). At a given proton flux, decreasing g(H)(+) will increase transthylakoid proton motive force (pmf), thus lowering lumen pH and contributing to the activation of NPQ. It was found that an approximately 5-fold decrease in g(H)(+) could account for the majority of NPQ modulation as atmospheric CO(2) was decreased from 2,000 ppm to 0 ppm. Data are presented that g(H)(+) is kinetically controlled, rather than imposed thermodynamically by buildup of DeltaG(ATP). Further results suggest that the redox state of the ATP synthase gamma-subunit thiols is not responsible for altering g(H)(+). A working model is proposed wherein g(H)(+) is modulated by stromal metabolite levels, possibly by inorganic phosphate.

Arabidopsis↗
PubMed2001

Definition of EGF-like, closely interacting modules that bear activation epitopes in integrin beta subunits.

Integrin beta subunits contain four cysteine-rich repeats in a long extracellular stalk that connects the headpiece to the membrane. Most mAbs to integrin activation epitopes map to these repeats, and they are important in propagating conformational signals from the membrane/cytosol to the ligand-binding headpiece. Sequence analysis of a protein containing only 10 integrin-like, cysteine-rich repeats suggests that these repeats start one cysteine earlier than previously reported. By using the new repeat boundaries, statistically significant sequence homology to epidermal growth factor-like domains is found, and a disulfide bond connectivity of the eight cysteines is predicted that differs in three of four disulfides from a previous prediction of epidermal growth factor-like modules [Berg, R. W., Leung, E., Gough, S., Morris, C., Yao, W.-P., Wang, S.-x., Ni, J. & Krissansen, G. W. (1999) Genomics 56, 169-178]. N-terminally truncated beta2 integrin stalk fragments were well expressed and secreted from 293 T cells when they began at repeat boundaries but not when they began one cysteine earlier or later. Furthermore, peptides that correspond to module 3 or modules 2 + 3 were expressed in bacteria and refolded. The module 2 + 3 fragment was as reactive with three mAbs to activation epitopes as a beta2 fragment expressed in eukaryotic cells, indicating a native fold. Only one residue intervenes between the last cysteine of one module and the first cysteine of the next. This arrangement is consistent with a tight intermodule connection, a prerequisite for signal propagation from the membrane to the ligand binding headpiece.

Amino Acid Sequence↗
PubMed1981

Modulation of Schistosoma mansoni egg-induced granuloma formation: I-J restriction of T cell-mediated suppression in a chronic parasitic infection.

Newly formed hepatic granulomas around Schistosoma mansoni eggs become progressively smaller during the chronic (greater than or equal to 15 weeks after infection) phase of the disease. This reduction in granuloma size, termed "modulation," is known to be caused in part by a T lymphocyte that can adoptively transfer modulation to 6-week-infected mice. The present study examines a possible role for the I-J locus in regulating the suppressor T lymphocyte aspects of modulation. Adoptive transfer between congeneic B10.A(3R) and B10.A(5R) mice (differing at the I-J locus) indicated that optimal suppression is dependent upon homology at the I-J locus. In vivo treatment of chronically infected mice with microliter amounts of antiserum specific for the recipient's I-J determinant blocked modulation during chronic infection and prevented adoptive transfer of suppression to 6-week-infected mice. The in vivo regimen of anti-I-J had no effect on anti-schistosomal egg antigen titers during chronic infection. These results demonstrate an I-J restriction for suppression. It appears that the suppressor T lymphocyte circuit responsible for this aspect of modulation requires an I-J positive lymphocyte.

Animals↗
PubMed1982

Modulation of T-lymphocyte differentiation antigens: potential relevance for multiple sclerosis.

Effects of the anti-T-cell monoclonal antibodies OKT3, OKT5, and OKT8 on T-cell surface properties and cell functions were evaluated. Incubation of mononuclear cells isolated from peripheral blood for 48 hr with each monoclonal antibody in the absence of complement resulted in modulation of their respective surface antigens; i.e., the number of cells detected by immunofluorescence as positive for the T3, T5, and T8 surface antigens was reduced. T3, T5, and T8 antigens modulated independently. A radiolabeled second antibody technique confirmed modulation by OKT3 and OKT8 and indicated that T-cell differentiation antigens can regenerate in culture. Incubation of mononuclear cells with OKT3 increased the number of sheep erythrocyte-binding lymphocytes (E+-rosetting cells) and markedly increased the number of avidly E+-rosetting cells. Incubation with OKT8 reduced the number of E+- and of avidly E+-rosetting cells. OKT3 induced both mitogenic reactivity and suppressor cell activity; cells modulated by OKT8 exhibited reduced mitogenic reactivity and reduced suppressor cell function. The decreases in total T cells, in avid T cells, in suppressor cell number, and in suppressor cell function that follow modulation by OKT8 mimic changes observed in multiple sclerosis patients.

Adult↗
PubMed1989

Cis-acting elements of the sea urchin histone H2A modulator bind transcriptional factors.

Functional tests, performed by microinjection into Xenopus laevis oocytes, show that a DNA fragment containing the modulator of the early histone H2A gene of Paracentrotus lividus enhances transcription of a reporter gene when located, in the physiological orientation, upstream of the tk basal promoter. Gel retardation and DNase I footprinting assays further reveal that the H2A modulator contains at least two binding sites [upstream sequence elements 1 and 2 (USE 1 and USE 2)] for nuclear factors extracted from sea urchin embryos, which actively transcribe the early histone gene set. Interestingly, USE 1 is highly homologous to a cis-acting element previously identified in the H2A modulator of Psammechinus miliaris [Grosschedl, R., Mächler, M., Rohrer, U. & Birnstiel, M. L. (1983) Nucleic Acids Res. 11, 8123-8136]. Finally, a cloned oligonucleotide containing the USE 1 sequence competes efficiently in Xenopus oocytes with the H2A modulator to prevent enhancement of transcription of the reporter gene. From these results, we conclude that USE 1 and perhaps USE 2 in the H2A modulator are upstream transcriptional elements that are recognized by trans-acting factors common to Xenopus and sea urchin.

Animals↗
PubMed1993

Crosstalk between alpha/beta T cells and gamma/delta T cells in vivo: activation of alpha/beta T-cell responses after gamma/delta T-cell modulation with the monoclonal antibody GL3.

Although gamma/delta T cells express numerous in vitro functions similar to alpha/beta T cells, little is known about their biological functioning in vivo. Furthermore, it is unclear whether alpha/beta T cells and gamma/delta T cells act independently or in a coordinated way. In the present study, gamma/delta T cells were modulated in vivo by i.p. injection of the anti-gamma/delta T-cell receptor (TCR) monoclonal antibody GL3. GL3 administration caused disappearance of the gamma/delta TCR in spleen and lymph node cells and the gamma/delta TCR was reexpressed after in vitro cultivation for a few days. When cultured in vitro for 4 days, in the absence of foreign antigens, spleen and lymph node alpha/beta T cells from GL3-modulated mice showed vigorous proliferative responses. CD4 T lymphocytes from GL3-modulated mice produced interleukin 2, and CD8 T cells developed into cytolytic T lymphocytes in vitro capable of lysing syngeneic and allogeneic targets. Treatment with heat-inactivated GL3 or with normal hamster immunoglobulin did not cause any of these effects. These findings suggest that the anti-gamma/delta TCR monoclonal antibody GL3 modulates gamma/delta T cells in vivo and that this modulation has profound effects on alpha/beta T-cell reactivity. Hence, the data suggest a role for gamma/delta T cells in the regulation of alpha/beta T-cell activation in vivo.

Animals↗
PubMed1995

Iontophoresis for modulation of cardiac drug delivery in dogs.

Cardiac arrhythmias are a frequent cause of death and morbidity. Conventional antiarrhythmia therapy involving oral or intravenous medication is often ineffective and complicated by drug-associated side effects. Previous studies from our laboratory have demonstrated the advantages of cardiac drug-polymer implants for enhanced efficacy for cardiac arrhythmia therapy compared with conventional administration. However, these studies were based on systems that deliver drugs at a fixed release rate. Modulation of the drug delivery rate has the advantage of regulating the amount of the drug delivered depending upon the disease state of the patient. We hypothesized that iontophoresis could be used to modulate cardiac drug delivery. In this study, we report our investigations of a cardiac drug implant in dogs that is capable of iontophoretic modulation of the administration of the antiarrhythmic agent sotalol. We used a heterogeneous cation-exchange membrane (HCM) as an electrically sensitive and highly efficient rate-limiting barrier on the cardiac-contacting surface of the implant. Thus, electric current is passed only through the HCM and not the myocardium. The iontophoretic cardiac implant demonstrated in vitro drug release rates that were responsive to current modulation. In vivo results in dogs have confirmed that iontophoresis resulted in regional coronary enhancement of sotalol levels with current-responsive increases in drug concentrations. We also observed acute current-dependent changes in ventricular effective refractory periods reflecting sotalol-induced refractoriness due to regional drug administration. In 30-day dog experiments, iontophoretic cardiac implants demonstrated robust sustained function and reproducible modulation of drug delivery kinetics.

Animals↗
PubMed1999

The curvature of a Hilbert module over.

A notion of curvature is introduced in multivariable operator theory. The curvature invariant of a Hilbert module over C[z(1),., z(d)] is a nonnegative real number which has significant extremal properties, which tends to be an integer, and which is hard to compute directly. It is shown that for graded Hilbert modules, the curvature agrees with the Euler characteristic of a certain finitely generated algebraic module over the appropriate polynomial ring. This result is a higher dimensional operator-theoretic counterpart of the Gauss-Bonnet formula which expresses the average Gaussian curvature of a compact oriented Riemann surface as the alternating sum of the Betti numbers of the surface, and it solves the problem of calculating the curvature of graded Hilbert modules. The proof of that result is based on an asymptotic formula which expresses the curvature of a Hilbert module in terms that allow its comparison to a corresponding asymptotic expression for the Euler characteristic.

Journal Article↗
PubMed1995

Alpha-scorpion toxins binding on rat brain and insect sodium channels reveal divergent allosteric modulations by brevetoxin and veratridine.

At least six topologically separated neurotoxin receptor sites have been identified on sodium channels that reveal strong allosteric interactions among them. We have studied the allosteric modulation induced by veratridine, binding to receptor site 2, and brevetoxin PbTx-1, occupying receptor site 5, on the binding of alpha-scorpion toxins at receptor site 3, on three different neuronal sodium channels: rat brain, locust, and cockroach synaptosomes. We used 125I-AaH II, the most active alpha-scorpion toxin on vertebrates, and 125I-Lqh alpha IT, shown to have high activity on insects, as specific probes for receptor site 3 in rat brain and insect sodium channels. Our results reveal that brevetoxin PbTx-1 generates three types of effects at receptor site 3:1) negative allosteric modulation in rat brain sodium channels, 2) positive modulation in locust sodium channels, and 3) no effect on cockroach sodium channel. However, PbTx-1 activates sodium channels in cockroach axon similarly to its activity in other preparation. Veratridine positively modulates both rat brain and locust sodium channels but had no effect on alpha-toxin binding in cockroach. The dramatic differences in allosteric modulations in each sodium channel subtype suggest structural differences in receptor sites for PbTx-1 and/or at the coupling regions with alpha-scorpion toxin receptor sites in the different sodium channels, which can be detected by combined application of specific channel modifiers and may elucidate the dynamic gating activity and the mechanism of allosteric interactions among various neurotoxin receptors.

Allosteric Regulation↗
PubMed1997

Disruption of ribosomal scanning on the 5'-untranslated region, and not restriction of translational initiation per se, modulates the stability of nonaberrant mRNAs in the yeast Saccharomyces cerevisiae.

Translation and mRNA decay constitute key players in the post-transcriptional control of gene expression. We examine the mechanisms by which the 5'-untranslated region (UTR) of nonaberrant mRNAs acts to modulate both these processes in Saccharomyces cerevisiae. Two classes of functional relationship between ribosome-5'-UTR interactions and mRNA decay are identifiable. In the first of these, elements in the main open reading frame (ORF) dictate how the decay process reacts to inhibitory structures in the 5'-UTR. The same types of stability modulation can be elicited by trans-regulation of translation via inducible binding of the iron-regulatory protein to an iron-responsive element located 9 nucleotides from the 5' cap. A eukaryotic translational repressor can therefore modulate mRNA decay via the 5'-UTR. In contrast, translational regulation mediated via changes in the activity of the cap-binding eukaryotic translation initiation factor eIF-4E bypasses translation-dependent pathways of mRNA degradation. Thus modulation of mRNA stability via the 5'-UTR depends on disruption of the scanning process, rather than changes in translational initiation efficiency per se. In the second class of pathway, an upstream ORF (uORF) functions as a powerful destabilizing element, inducing termination-dependent degradation that is apparently independent of any main ORF determinants but influenced by the efficiencies of ribosomal recognition of the uORF start and stop codons. This latter mechanism provides a regulatable means to modulate the stability of nonaberrant mRNAs via a UPF-dependent pathway.

Eukaryotic Initiation Factor-4E↗
PubMed1997

Interaction of the NH2-terminal domain of fibronectin with heparin. Role of the omega-loops of the type I modules.

Determinants of the interaction of the 29-kDa NH2-terminal domain of fibronectin with heparin were explored by analysis of normal and mutant recombinant NH2-terminal fibronectin fragments produced in an insect cell Baculovirus host vector system. A genomic/cDNA clone was constructed that specified a secretable human fibronectin NH2 fragment. With the use of site-directed mutagenesis a set of 29 kDa fragments was obtained that contained glycine or glutamic acid residues in place of basic residues at various candidate sites for heparin binding in the five type I modules that make up the domain. The recombinant fragment containing the wild type sequence had a nearly normal circular dichroic spectra and a melting profile, as assayed by loss of ellipticity at 228 nm, that was indistinguishable from that of the native fragment obtained by trypsinization of plasma fibronectin. A substantial proportion of the wild type recombinant fragment bound to heparin-Sepharose, where it was eluted at the same NaCl concentration as the native fragment. The wild type fragment was capable of promoting matrix-driven translocation, a morphogenetic effect in artificial extracellular matrices that depends on the interaction of the fibronectin NH2 terminus with heparin-like molecules on the surfaces of particles. Mutant fragments in which arginines predicted to be most exposed in the folded fragment were converted to glycines retained the same affinity for heparin as the wild type fragment. In contrast, a mutant fragment in which the single basic residue (Arg99) in the minor loop ("Omega-loop") of the second type I module was converted to a glycine had an essentially normal melting profile but exhibited no binding to heparin and failed to promote matrix-driven translocation. A mutant fragment in which the single basic residue (Arg52) of the first type I module was converted to a glycine also completely lacked heparin binding activity, but one in which the single basic residue (Arg191) the fourth type I module was converted to a glycine retained the ability to bind heparin. A mutant fragment in which the single basic residue (Lys143) in the Omega-loop of the third type I module was converted to a glutamic acid lacked heparin binding activity but had a CD spectrum similar to the heparin-liganded native protein and was capable of promoting matrix-driven translocation. The results indicate that multiple residues in the Omega-loops of the fibronectin NH2-terminal domain participate in its interactions with heparin. In addition, the conformation of one of the nonbinding mutants may mimic the heparin-induced structural alteration in this fibronectin domain required for certain morphogenetic events.

Amino Acid Sequence↗
PubMed1998

Baculovirus-mediated expression of truncated modular fragments from the catalytic region of human complement serine protease C1s. Evidence for the involvement of both complement control protein modules in the recognition of the C4 protein substrate.

C1s is the modular serine protease responsible for cleavage of C4 and C2, the protein substrates of the first component of complement. Its catalytic region (gamma-B) comprises two complement control protein (CCP) modules, a short activation peptide (ap), and a serine protease domain (SP). A baculovirus-mediated expression system was used to produce recombinant truncated fragments from this region, deleted either from the first CCP module (CCP2-ap-SP) or from both CCP modules (ap-SP). The aglycosylated fragment CCP2-ap-SPag was also expressed by using tunicamycin. The fragments were produced at yields of 0.6-3 mg/liter of culture, isolated, and characterized chemically and then tested functionally by comparison with intact C1s and its proteolytic gamma-B fragment. All recombinant fragments were expressed in a proenzyme form and cleaved by C1r to generate active enzymes expressing esterolytic activity and reactivity toward C1 inhibitor comparable to those of intact C1s. Likewise, the activated fragments gamma-B, CCP2-ap-SP, and ap-SP retained C1s ability to cleave C2 in the fluid phase. In contrast, whereas fragment gamma-B cleaved C4 as efficiently as C1s, the C4-cleaving activity of CCP2-ap-SP was greatly reduced (about 70-fold) and that of ap-SP was abolished. It is concluded that C4 cleavage involves substrate recognition sites located in both CCP modules of C1s, whereas C2 cleavage is affected mainly by the serine protease domain. Evidence is also provided that the carbohydrate moiety linked to the second CCP module of C1s has no significant effect on catalytic activity.

Animals↗