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151.
Interaction of fibrin(ogen) with heparin: further characterization and localization of the heparin-binding site 总被引:1,自引:0,他引:1
The beta chain 15-42 sequence of the fibrin(ogen) E region was implicated in heparin binding [Odrljin et al. (1996) Blood 88, 2050-2061]; whether heparin binds to other fibrin(ogen) regions remains to be clarified. To address this question, we studied the interaction of heparin with fibrinogen, fibrin, and their major fragments D(1), D-D, E(1), E(3), and alphaC, which together cover the entire structure of the molecule, by ligand blotting, surface plasmon resonance, and fluorescence. All three techniques revealed that at physiological ionic conditions only fibrin(ogen) and the E(1) fragment bind heparin, indicating that the only physiologically relevant heparin-binding site of fibrin(ogen) is located in its E region. To test whether the beta15-42 sequence is sufficient to form this site or some additional sequences are also involved, we tested the interaction of heparin with a number of beta15-42-containing fragments. The synthetic beta15-42 peptide bound heparin weakly (K(d) = 44.5 microM) while the recombinant beta15-57 and beta15-64 fragments exhibited almost 7-fold higher affinity (K(d) = 6.4 and 7.1 microM, respectively), indicating that the beta43-57 region is also important for heparin binding. At the same time the recombinant dimeric disulfide-linked (beta15-66)(2) fragment which mimics the dimeric arrangement of the beta chains in fibrin bound heparin with high affinity (K(d) = 66 nM), almost 100-fold higher than that for the monomeric fragments. This affinity was similar to those determined for fibrin and the E(1) fragment (K(d) = 72 and 70 nM, respectively) suggesting that (beta15-66)(2) mimics well the heparin-binding properties of the latter two. Altogether, these results indicate that the only heparin-binding site in fibrin(ogen) is formed by NH(2)-terminal portions of the beta chains, including residues beta15-57, and that dimerization is essential for high-affinity binding. 相似文献
152.
Yevdokimova NY Veselovska LD Gogolinska GK Buchanevich OM Kosyakova GV Gritsenko PG Lugovskoj EV Komisarenko SV 《Acta biochimica Polonica》2003,50(1):279-289
Fibrin split product D-dimer (DD) is most probably involved in the development of vascular disorders. At 1.5 microM concentration DD inhibited the incorporation of D-[1-(3)H]glucosamine hydrochloride and [2-(14)C]acetate x Na into pericellular heparan sulphate (HS) of rabbit coronary endothelial cells without affecting other groups of glycosaminoglycans (GAGs). At the same time, DD reduced HS ability to bind antithrombin (AT) and suppressed NO production. The effect of DD on pericellular GAGs was similar to that of N(omega)-methyl-L-arginine, the competitive inhibitor of endothelial NO synthase (eNOS). L-Ascorbic acid, eNOS activator, increased the level of endogenous NO in the DD-treated cells, and restored HS accumulation and antithrombin binding. It is suggested that DD influence on endothelial HS may be mediated by NO production. Another effect of DD, namely, stimulation of plasminogen activator inhibitor-1 (PAI-1) secretion did not depend on the NO level. The decreased HS content, reduced anticoagulant properties of HS, and increased PAI-1 secretion disorganized the endothelial matrix, and promoted fibrin formation and vascular damage. This points to DD as an important factor in the development of vascular disorders. 相似文献
153.
Ectopic expression of atRSZ33 reveals its function in splicing and causes pleiotropic changes in development 下载免费PDF全文
Splicing provides an additional level in the regulation of gene expression and contributes to proteome diversity. Herein, we report the functional characterization of a recently described plant-specific protein, atRSZ33, which has characteristic features of a serine/arginine-rich protein and the ability to interact with other splicing factors, implying that this protein might be involved in constitutive and/or alternative splicing. Overexpression of atRSZ33 leads to alteration of splicing patterns of atSRp30 and atSRp34/SR1, indicating that atRSZ33 is indeed a splicing factor. Moreover, atRSZ33 is a regulator of its own expression, as splicing of its pre-mRNA is changed in transgenic plants. Investigations by promoter-beta-glucuronidase (GUS) fusion and in situ hybridization revealed that atRSZ33 is expressed during embryogenesis and early stages of seedling formation, as well as in flower and root development. Ectopic expression of atRSZ33 caused pleiotropic changes in plant development resulting in increased cell expansion and changed polarization of cell elongation and division. In addition, changes in activity of an auxin-responsive promoter suggest that auxin signaling is disturbed in these transgenic plants. 相似文献
154.
We demonstrate that chain length is the main determinant of the folding rate for proteins with the three-state folding kinetics. The logarithm of their folding rate in water (k(f)) strongly anticorrelates with their chain length L (the correlation coefficient being -0.80). At the same time, the chain length has no correlation with the folding rate for two-state folding proteins (the correlation coefficient is -0.07). Another significant difference of these two groups of proteins is a strong anticorrelation between the folding rate and Baker's "relative contact order" for the two-state folders and the complete absence of such correlation for the three-state folders. 相似文献
155.
Blind noise reduction for multisensory signals using ICA and subspace filtering, with application to EEG analysis 总被引:8,自引:0,他引:8
In many applications of signal processing, especially in communications and biomedicine, preprocessing is necessary to remove
noise from data recorded by multiple sensors. Typically, each sensor or electrode measures the noisy mixture of original source
signals. In this paper a noise reduction technique using independent component analysis (ICA) and subspace filtering is presented.
In this approach we apply subspace filtering not to the observed raw data but to a demixed version of these data obtained
by ICA. Finite impulse response filters are employed whose vectors are parameters estimated based on signal subspace extraction.
ICA allows us to filter independent components. After the noise is removed we reconstruct the enhanced independent components
to obtain clean original signals; i.e., we project the data to sensor level. Simulations as well as real application results
for EEG-signal noise elimination are included to show the validity and effectiveness of the proposed approach.
Received: 6 November 2000 / Accepted in revised form: 12 November 2001 相似文献
156.
157.
Leytin V Allen DJ Mykhaylov S Mis L Lyubimov EV Garvey B Freedman J 《Biochemical and biophysical research communications》2004,320(2):303-310
Recently, it has been discovered that apoptosis of anucleate platelets can be induced by chemical agonists. Other studies demonstrated that mechanical forces (shear stresses) stimulate platelet activation and signaling in the absence of exogenous chemical stimuli. We analyzed whether shear stresses can trigger platelet apoptosis, a question that has not yet been studied. Using a cone-and-plate viscometer, we exposed human platelet-rich plasma to different shear stresses, ranging from physiologic arterial and arteriole levels (10-44 dyn/cm2) to pathologic high levels (117-388 dyn/cm2) occurring in stenotic vessels. We found that pathologic shear stresses induce not only platelet activation (P-selectin upregulation and GPIbalpha downregulation) but also trigger apoptosis events, including mitochondrial transmembrane potential depolarization, caspase 3 activation, phosphatidylserine exposure, and platelet shrinkage and fragmentation, whereas physiological shear stresses are not effective.This novel finding suggests that shear-induced platelet apoptosis can be mediated by mechanoreceptors, does not require nuclear participation, and may affect platelet clearance. 相似文献
158.
Our theoretical approach for prediction of folding/unfolding nuclei in three-dimensional protein structures is based on a search for free energy saddle points on networks of protein unfolding pathways. Under some approximations, this search is performed rapidly by dynamic programming and results in prediction of Phi values, which can be compared with those found experimentally. In this study, we optimize some details of the model (specifically, hydrogen atoms are taken into account in addition to heavy atoms), and compare the theoretically obtained and experimental Phi values (which characterize involvement of residues in folding nuclei) for all 17 proteins, where Phi values are now known for many residues. We show that the model provides good Phi value predictions for proteins whose structures have been determined by X-ray analysis (the average correlation coefficient is 0.65), with a more limited success for proteins whose structures have been determined by NMR techniques only (the average correlation coefficient is 0.34), and that the transition state free energies computed from the same model are in a good anticorrelation with logarithms of experimentally measured folding rates at mid-transition (the correlation coefficient is -0.73). 相似文献
159.
160.
Commercially available high-content Ab380 and extensively validated DLM26 homemade protein microarrays were used to profile the effects of the pro-atherogenic molecule, oxidized low density lipoprotein (OxLDL), on human aortic smooth muscle cells. Protein microarrays detected 298 proteins in cell lysates and 54 of these were differentially regulated. Microarray data were validated by immunoblotting for a selected set of up- and down-regulated proteins. The protein microarray data sets were compared with our recent cDNA microarray-based gene expression results in order to characterize the global effect of OxLDL on smooth muscle cell functions. A group of cell-cell interaction molecules was classified as up-regulated by OxLDL, whereas nucleic acid/protein biosynthesis, structural and humoral response proteins/genes were under-expressed in cells treated by OxLDL. These findings reveal the major pattern of OxLDL-induced effects on the human aortic smooth muscle cells functions and also demonstrate that protein chip-based microarrays could be a useful proteomic tool to profile disease-related states of muscle cells. 相似文献