FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2066231048> ?p ?o ?g. }

• W2066231048 endingPage "15586" @default.
• W2066231048 startingPage "15583" @default.
• W2066231048 abstract "The endothelial nitric-oxide synthase (eNOS) is a key determinant of vascular homeostasis. Like all known nitric-oxide synthases, eNOS enzyme activity is dependent on Ca2+-calmodulin. eNOS is dynamically targeted to specialized cell surface signal-transducing domains termed plasmalemmal caveolae and interacts with caveolin, an integral membrane protein that comprises a key structural component of caveolae. We have previously reported that the association between eNOS and caveolin is quantitative and tissue-specific (Feron, O., Belhassen, L., Kobzick, L., Smith, T. W., Kelly, R. A., and Michel, T. (1996) J. Biol. Chem.271, 22810–22814). We now report that in endothelial cells the interaction between eNOS and caveolin is importantly regulated by Ca2+-calmodulin. Addition of calmodulin disrupts the heteromeric complex formed between eNOS and caveolin in a Ca2+-dependent fashion. In addition, overexpression of caveolin markedly attenuates eNOS enzyme activity, but this inhibition is reversed by purified calmodulin. Caveolin overexpression does not affect the activity of the other NOS isoforms, suggesting eNOS-specific inhibition of NO synthase by caveolin. We propose a model of reciprocal regulation of eNOS in endothelial cells wherein the inhibitory eNOS-caveolin complex is disrupted by binding of Ca2+-calmodulin to eNOS, leading to enzyme activation. These findings may have broad implications for the regulation of Ca2+-dependent signal transduction in plasmalemmal caveolae. The endothelial nitric-oxide synthase (eNOS) is a key determinant of vascular homeostasis. Like all known nitric-oxide synthases, eNOS enzyme activity is dependent on Ca2+-calmodulin. eNOS is dynamically targeted to specialized cell surface signal-transducing domains termed plasmalemmal caveolae and interacts with caveolin, an integral membrane protein that comprises a key structural component of caveolae. We have previously reported that the association between eNOS and caveolin is quantitative and tissue-specific (Feron, O., Belhassen, L., Kobzick, L., Smith, T. W., Kelly, R. A., and Michel, T. (1996) J. Biol. Chem.271, 22810–22814). We now report that in endothelial cells the interaction between eNOS and caveolin is importantly regulated by Ca2+-calmodulin. Addition of calmodulin disrupts the heteromeric complex formed between eNOS and caveolin in a Ca2+-dependent fashion. In addition, overexpression of caveolin markedly attenuates eNOS enzyme activity, but this inhibition is reversed by purified calmodulin. Caveolin overexpression does not affect the activity of the other NOS isoforms, suggesting eNOS-specific inhibition of NO synthase by caveolin. We propose a model of reciprocal regulation of eNOS in endothelial cells wherein the inhibitory eNOS-caveolin complex is disrupted by binding of Ca2+-calmodulin to eNOS, leading to enzyme activation. These findings may have broad implications for the regulation of Ca2+-dependent signal transduction in plasmalemmal caveolae. Nitric oxide is a ubiquitous molecule implicated in diverse biological processes and is synthesized in mammalian cells by a family of Ca2+-calmodulin-dependent nitric-oxide synthase (NOS) 1The abbreviations used are: NOS, nitric-oxide synthase; NO, nitric oxide; eNOS, endothelial isoform of NOS; iNOS, inducible isoform of NOS; nNOS, neuronal isoform of NOS; PAGE, polyacrylamide gel electrophoresis; CHAPS, 3-[(3-holamidopropyl)dimethylammonio]-1-propanesulfonic acid. enzymes (1Nathan C. Xie Q.-W. J. Biol. Chem. 1994; 269: 13725-13728Abstract Full Text PDF PubMed Google Scholar, 2Marletta M. Cell. 1994; 78: 927-930Abstract Full Text PDF PubMed Scopus (817) Google Scholar, 3Sase K. Michel T Trends Cardiovasc. Med. 1997; 7: 28-37Crossref PubMed Scopus (84) Google Scholar). Endothelium-derived nitric oxide (NO), formed by the endothelial isoform of nitric-oxide synthase (eNOS), serves as an important determinant of blood pressure and platelet aggregation. In endothelial cells, increases in intracellular Ca2+ elicited by diverse extracellular signals lead to activation of eNOS. The three known mammalian nitric-oxide synthases share similar overall Ca2+-calmodulin-dependent catalytic pathways. However, the eNOS enzyme is unique among the three known NOS isoforms in being localized to the specialized cell surface signal-transducing domains termed plasmalemmal caveolae (4Shaul P.W. Smart E.J. Robinson L.J. German Z. Yuhanna I.S. Ying Y. Anderson R.G.W. Michel T. J. Biol. Chem. 1996; 271: 6518-6522Abstract Full Text Full Text PDF PubMed Scopus (626) Google Scholar, 5Feron O. Belhassen L. Kobzick L. Smith T.W. Kelly R.A. Michel T. J. Biol. Chem. 1996; 271: 22810-22814Abstract Full Text Full Text PDF PubMed Scopus (598) Google Scholar). Plasmalemmal caveolae are small invaginations in the plasma membrane that may serve as sites for the sequestration of signaling proteins (6Anderson R.G. Proc. Natl. Acad. Sci. U. S. A. 1993; 90: 10909-10913Crossref PubMed Scopus (570) Google Scholar,7Lisanti M.P. Scherer P.E. Vidugiriene J. Tang Z. Hermanowski-Vosatka A. Tu Y.H. Cook R.F. Sargiacomo M. J. Cell. Biol. 1995; 126: 111-126Crossref Scopus (815) Google Scholar) including receptors, G proteins, and protein kinases, as well as eNOS. The principal protein in caveolae is the integral membrane protein caveolin, an oligomeric protein that serves as a structural “scaffold” within caveolae (8Sargiacomo M. Scherer P.E. Tang Z.-L. Kubler E. Song K.S. Sanders M.C. Lisanti M.P. Proc. Natl. Acad. Sci. U. S. A. 1995; 92: 9407-9411Crossref PubMed Scopus (480) Google Scholar). eNOS can be quantitatively immunoprecipitated by antibodies directed against caveolin; conversely, eNOS antiserum also immunoprecipitates caveolin (5Feron O. Belhassen L. Kobzick L. Smith T.W. Kelly R.A. Michel T. J. Biol. Chem. 1996; 271: 22810-22814Abstract Full Text Full Text PDF PubMed Scopus (598) Google Scholar), although it has not yet been established whether the interaction between these two proteins is direct. Moreover, a functional role of the eNOS-caveolin association, beyond its postulated role in subcellular targeting of the enzyme, remains to be determined. We document in this report that the interaction between eNOS and caveolin may be regulated by Ca2+-calmodulin, and we show that caveolin can specifically inhibit eNOS enzyme activity. A plasmid construct encoding eNOS has been described previously (9Busconi L. Michel T. J. Biol. Chem. 1993; 268: 8410-8413Abstract Full Text PDF PubMed Google Scholar, 10Lamas S. Marsden P.A. Li G. Tempst P. Michel T. Proc. Natl. Acad. Sci. U. S. A. 1992; 89: 6348-6352Crossref PubMed Scopus (921) Google Scholar). cDNA clones encoding iNOS (11Xie Q.-W. Cho H.J. Calaycay J. Mumford R.A. Swiderek K.M. Lee T.M. Ding A. Troso T. Nathan C. Science. 1992; 256: 225-228Crossref PubMed Scopus (1741) Google Scholar) and nNOS (12Bredt D.S. Hwang P.M. Glatt C.E. Lowenstein C. Reed R.R. Snyder S.H. Nature. 1991; 351: 714-718Crossref PubMed Scopus (2173) Google Scholar) were kindly provided by Carl Nathan (Cornell University Medical College) and Solomon Snyder (Johns Hopkins University), respectively. Caveolin-1 cDNA (13Li S. Couet J. Lisanti M.P. J. Biol. Chem. 1996; 271: 29182-29190Abstract Full Text Full Text PDF PubMed Scopus (675) Google Scholar) in the eukaryotic expression vector pCB-7 was obtained from Michael Lisanti (Whitehead Institute). An irrelevant plasmid encoding β-galactosidase was used as a control to maintain identical amounts of DNA in each transfection. Cultures of bovine aortic endothelial cells (studied between passages 4 and 10) and COS-7 cells were performed as described previously (9Busconi L. Michel T. J. Biol. Chem. 1993; 268: 8410-8413Abstract Full Text PDF PubMed Google Scholar, 14Lee C. Robinson L.J. Michel T. J. Biol. Chem. 1995; 270: 27403-27406Abstract Full Text Full Text PDF PubMed Scopus (57) Google Scholar). COS-7 cells were transfected with 10 μg of total plasmid DNA in 100-mm cell culture plates using LipofectAMINE™ (Life Technologies, Inc.) according to the manufacturer's protocol. Endothelial cells were lysed and solubilized either with: 1) a previously described Ca2+-free CHAPS buffer (5Feron O. Belhassen L. Kobzick L. Smith T.W. Kelly R.A. Michel T. J. Biol. Chem. 1996; 271: 22810-22814Abstract Full Text Full Text PDF PubMed Scopus (598) Google Scholar, 14Lee C. Robinson L.J. Michel T. J. Biol. Chem. 1995; 270: 27403-27406Abstract Full Text Full Text PDF PubMed Scopus (57) Google Scholar) supplemented with 1 mm EGTA/1 mmEDTA or 2) an otherwise identical CHAPS buffer containing 1 mm CaCl2 and no EDTA/EGTA. CHAPS-solubilized bovine aortic endothelial cell lysates were incubated either with a polyclonal caveolin antibody (Transduction Laboratories) at a final concentration of 4 μg/ml or with a previously characterized polyclonal antiserum directed against eNOS (9Busconi L. Michel T. J. Biol. Chem. 1993; 268: 8410-8413Abstract Full Text PDF PubMed Google Scholar) used at a final dilution of 1:100. Immunoprecipitated complexes were then recovered, denatured in Laemmli sample buffer, separated on 12% SDS-PAGE, and transferred to a polyvinylidene difluoride membrane (5Feron O. Belhassen L. Kobzick L. Smith T.W. Kelly R.A. Michel T. J. Biol. Chem. 1996; 271: 22810-22814Abstract Full Text Full Text PDF PubMed Scopus (598) Google Scholar). Monoclonal antibodies directed against eNOS or caveolin-1 (Transduction Laboratories) were then used to detect eNOS and caveolin-1 using chemiluminescence, as described previously (5Feron O. Belhassen L. Kobzick L. Smith T.W. Kelly R.A. Michel T. J. Biol. Chem. 1996; 271: 22810-22814Abstract Full Text Full Text PDF PubMed Scopus (598) Google Scholar). Calmodulin was detected using a previously characterized monoclonal calmodulin antibody (15Sacks D.B. Porter S.E. Ladenson J.H. McDonald J.M. Anal. Biochem. 1991; 194: 369-377Crossref PubMed Scopus (79) Google Scholar). Expression of immunoblotted proteins was quantitated by laser densitometric analysis of x-ray films following chemiluminescence detection. NO synthase activity in lysates prepared from transfected COS-7 cell was determined by measuring conversion of [3H]l-arginine to [3H]l-citrulline as described previously (9Busconi L. Michel T. J. Biol. Chem. 1993; 268: 8410-8413Abstract Full Text PDF PubMed Google Scholar,14Lee C. Robinson L.J. Michel T. J. Biol. Chem. 1995; 270: 27403-27406Abstract Full Text Full Text PDF PubMed Scopus (57) Google Scholar). In some experiments, this NO synthase activity assay was performed in washed membrane fractions prepared from endothelial cell lysates as described previously (9Busconi L. Michel T. J. Biol. Chem. 1993; 268: 8410-8413Abstract Full Text PDF PubMed Google Scholar), except that calmodulin concentrations were varied by the addition of purified bovine brain calmodulin (Sigma) as indicated. In exploring the factors governing eNOS-caveolin association, we discovered that the co-immunoprecipitation of eNOS and caveolin was markedly attenuated when endothelial cell lysates were prepared in buffers containing excess free Ca2+ (Fig.1). Under no conditions did the presence of Ca2+ affect the recovery of caveolin or eNOS from endothelial cell lysates when the proteins were directlyimmunoprecipitated by their cognate antibody. Onlyco-immunoprecipitation was abrogated by Ca2+, as shown in Fig. 1. We next investigated the role of Ca2+ in modulating the association of eNOS and calmodulin. This is particularly important because calmodulin, a ubiquitous Ca2+-binding protein, plays a central role in nitric-oxide synthase catalysis (16Abu-Soud H.M. Stuehr D.J. Proc. Natl. Acad. Sci. U. S. A. 1992; 90: 10769-10772Crossref Scopus (398) Google Scholar). Co-immunoprecipitation experiments using eNOS antiserum to explore eNOS-calmodulin interactions in endothelial cell lysates are shown in Fig. 2. We could detect co-immunoprecipitation of eNOS and calmodulin only when free Ca2+ was present. This result is in striking contrast to the loss of eNOS-caveolin co-immunoprecipitation observed in the presence of Ca2+(Fig. 1). Taken together, these data suggest that Ca2+differentially modulates the association of eNOS with caveolinversus calmodulin in endothelial cell lysates. We next used antibodies against caveolin to co-immunoprecipitate eNOS in Ca2+-free buffers and then washed the immune complexes extensively to remove residual Ca2+ and calmodulin. Subsequent addition of either Ca2+ or calmodulin alone had no effect on the subsequent recovery of co-immunoprecipitated eNOS from the eNOS-caveolin complex (Fig. 3, upper panel). However, when Ca2+ and calmodulin were added together, eNOS was entirely lost from the caveolin immune complex. This “lost” eNOS could be completely recovered in the supernatant of the immune complex (Fig. 3, middle panel), indicating that the eNOS molecule had been released and not degraded following treatment of the caveolin-eNOS complex with Ca2+ plus calmodulin. None of these treatments affected the recovery of caveolin itself from the immune complex (Fig. 3, lower panel). The extensive washing of these immune complexes, required to remove the endogenous calmodulin, likely led to the loss of some eNOS (because the affinity of the eNOS-caveolin interaction is undoubtedly less than the affinity of the antibody for caveolin), leading to the detection of a relatively faint but highly reproducible (n = 4) signal for eNOS released from these complexes by the combination of Ca2+plus calmodulin. Diverse experimental approaches have shown that agonist activation of eNOS in endothelial cells is dependent on Ca2+-calmodulin (1Nathan C. Xie Q.-W. J. Biol. Chem. 1994; 269: 13725-13728Abstract Full Text PDF PubMed Google Scholar, 2Marletta M. Cell. 1994; 78: 927-930Abstract Full Text PDF PubMed Scopus (817) Google Scholar). The striking effects of Ca2+-calmodulin on the interactions of eNOS and caveolin therefore suggested to us that caveolin may influence eNOS enzyme activity. Indeed, caveolin has recently been shown in vitro to interact with other signaling proteins, including H-ras and G protein α subunits, preferentially associating with the “inactive” forms of these proteins (17Song K.S. Li S. Okamoto T. Quilliam L.A. Sargiacomo M. Lisanti M.P. J. Biol. Chem. 1996; 271: 9690-9697Abstract Full Text Full Text PDF PubMed Scopus (921) Google Scholar), but the cellular regulation of these interactions is less well understood. We explored the functional consequences of the interaction between eNOS and caveolin using transient transfection experiments in COS-7 cells and analyzed eNOS enzyme activity in transfected cells by assaying the conversion of [3H]l-arginine to [3H]l-citrulline in cell lysates, as shown in Fig. 4. In three separate experiments, each conducted in triplicate, we found that the co-transfection of a plasmid cDNA construct encoding caveolin with eNOS cDNA led to a marked attenuation of NOS activity (3.4 ± 0.3 versus 1.6 ± 0.1 pmol citrulline/min·mg protein in the absence or the presence of caveolin co-expression, respectively; see Fig. 4 A). There was no change in the abundance of eNOS protein associated with caveolin co-transfection, as assessed in immunoblots of these cellular lysates analyzed in each experiment (data not shown). Importantly, caveolin co-transfection failed to attenuate the enzyme activity expressed by transfected iNOS or nNOS cDNA, shown in Fig. 4 B. As for eNOS, the enzyme activity of iNOS and nNOS is calmodulin-dependent (although important differences in the Ca2+ dependence of the different NOS isoforms have been noted). In further contrast to eNOS, the other NOS isoforms are not targeted to caveolae. To explore the specificity of the inhibitory effect of caveolin co-expression on eNOS enzyme activity, we performed activity assays in the presence of varying concentrations of purified calmodulin added to washed membrane fractions prepared from transfected COS-7 cells. As shown in Fig. 4 C, in cells transfected with eNOS cDNA alone, there is a robust NOS activity even in the absence of added calmodulin (presumably due the presence of endogenous calmodulin in these membranes); enzyme activity increases only slightly with the addition of exogenous calmodulin. By contrast, caveolin co-expression markedly inhibits eNOS activity (by >90%) in the absence of added calmodulin; addition of increasing concentrations of exogenous calmodulin relieves this enzyme inhibition in a dose-dependent fashion, documenting that the caveolin inhibitory effect may be specifically overcome by purified calmodulin. Taken together, these studies suggest that the interaction between eNOS and caveolin is dynamically and specifically regulated by Ca2+-calmodulin and may serve as an important point of control in NO-dependent signaling. A direct interaction of caveolin with calmodulin appears unlikely to us because there was no influence of caveolin on the activity of other calmodulin-binding proteins (iNOS and nNOS) closely related to eNOS. This hypothesis is consistent with our failure to detect co-immunoprecipitation of calmodulin with caveolin (Fig. 2), under conditions in which eNOS was shown to associate with either one or the other protein. Furthermore, the amino acid sequence of caveolin isoforms show no obvious sequence or structural homologies to the known NOS isoforms (10Lamas S. Marsden P.A. Li G. Tempst P. Michel T. Proc. Natl. Acad. Sci. U. S. A. 1992; 89: 6348-6352Crossref PubMed Scopus (921) Google Scholar, 11Xie Q.-W. Cho H.J. Calaycay J. Mumford R.A. Swiderek K.M. Lee T.M. Ding A. Troso T. Nathan C. Science. 1992; 256: 225-228Crossref PubMed Scopus (1741) Google Scholar, 12Bredt D.S. Hwang P.M. Glatt C.E. Lowenstein C. Reed R.R. Snyder S.H. Nature. 1991; 351: 714-718Crossref PubMed Scopus (2173) Google Scholar) nor to any known calmodulin-binding protein sequences. Caveolin can attenuate the tyrosine kinase activity of c-src, an enzyme that bears no structural homology to eNOS, and is not known to be regulated by calmodulin (13Li S. Couet J. Lisanti M.P. J. Biol. Chem. 1996; 271: 29182-29190Abstract Full Text Full Text PDF PubMed Scopus (675) Google Scholar). We speculate that there is a common higher order structure assumed by the inactive conformation of diverse caveolae-targeted signaling proteins that forms the basis for their common interaction with caveolin. The targeting of eNOS to caveolae is likely to facilitate the interactions of eNOS with other co-localized signaling and regulatory molecules (3Sase K. Michel T Trends Cardiovasc. Med. 1997; 7: 28-37Crossref PubMed Scopus (84) Google Scholar). Formation of an inhibitory eNOS-caveolin heteromeric complex may serve to ensure the latency of the NO signal until calcium-mobilizing extracellular stimuli destabilize this complex and activate the enzyme. This close control of enzyme activity may be particularly important for eNOS in caveolae, where calmodulin, the enzyme's key allosteric activator, also is localized (4Shaul P.W. Smart E.J. Robinson L.J. German Z. Yuhanna I.S. Ying Y. Anderson R.G.W. Michel T. J. Biol. Chem. 1996; 271: 6518-6522Abstract Full Text Full Text PDF PubMed Scopus (626) Google Scholar) and where even subtle increases in intracellular Ca2+ could thus lead to enzyme activation if the interactions of caveolin with eNOS were not keeping the system in check. Because nitric oxide has cytotoxic as well as signaling functions (1Nathan C. Xie Q.-W. J. Biol. Chem. 1994; 269: 13725-13728Abstract Full Text PDF PubMed Google Scholar, 2Marletta M. Cell. 1994; 78: 927-930Abstract Full Text PDF PubMed Scopus (817) Google Scholar), attenuation of basal enzyme “leakiness” by caveolin may be of particular importance. The reciprocal regulation of eNOS by caveolin and calmodulin may represent a novel mechanism for the concerted control of NO production in the vascular wall. We are grateful to John Joyal for helpful discussions and for assistance performing calmodulin immunoblots." @default.
• W2066231048 created "2016-06-24" @default.
• W2066231048 creator A5012313192 @default.
• W2066231048 creator A5015223159 @default.
• W2066231048 creator A5059376263 @default.
• W2066231048 creator A5084285813 @default.
• W2066231048 date "1997-06-01" @default.
• W2066231048 modified "2023-10-17" @default.
• W2066231048 title "Reciprocal Regulation of Endothelial Nitric-oxide Synthase by Ca2+-Calmodulin and Caveolin" @default.
• W2066231048 cites W1507344018 @default.
• W2066231048 cites W1519147845 @default.
• W2066231048 cites W1549194476 @default.
• W2066231048 cites W1607802573 @default.
• W2066231048 cites W1995150314 @default.
• W2066231048 cites W1999565215 @default.
• W2066231048 cites W2003908597 @default.
• W2066231048 cites W2009013636 @default.
• W2066231048 cites W2023170094 @default.
• W2066231048 cites W2028880044 @default.
• W2066231048 cites W2033496106 @default.
• W2066231048 cites W2044918962 @default.
• W2066231048 cites W2068908631 @default.
• W2066231048 cites W2078331312 @default.
• W2066231048 cites W2084379296 @default.
• W2066231048 cites W2095480781 @default.
• W2066231048 cites W2144760773 @default.
• W2066231048 doi "https://doi.org/10.1074/jbc.272.25.15583" @default.
• W2066231048 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/9188442" @default.
• W2066231048 hasPublicationYear "1997" @default.
• W2066231048 type Work @default.
• W2066231048 sameAs 2066231048 @default.
• W2066231048 citedByCount "569" @default.
• W2066231048 countsByYear W20662310482012 @default.
• W2066231048 countsByYear W20662310482013 @default.
• W2066231048 countsByYear W20662310482014 @default.
• W2066231048 countsByYear W20662310482015 @default.
• W2066231048 countsByYear W20662310482016 @default.
• W2066231048 countsByYear W20662310482017 @default.
• W2066231048 countsByYear W20662310482018 @default.
• W2066231048 countsByYear W20662310482019 @default.
• W2066231048 countsByYear W20662310482020 @default.
• W2066231048 countsByYear W20662310482021 @default.
• W2066231048 countsByYear W20662310482022 @default.
• W2066231048 countsByYear W20662310482023 @default.
• W2066231048 crossrefType "journal-article" @default.
• W2066231048 hasAuthorship W2066231048A5012313192 @default.
• W2066231048 hasAuthorship W2066231048A5015223159 @default.
• W2066231048 hasAuthorship W2066231048A5059376263 @default.
• W2066231048 hasAuthorship W2066231048A5084285813 @default.
• W2066231048 hasBestOaLocation W20662310481 @default.
• W2066231048 hasConcept C112243037 @default.
• W2066231048 hasConcept C12554922 @default.
• W2066231048 hasConcept C138885662 @default.
• W2066231048 hasConcept C178790620 @default.
• W2066231048 hasConcept C181199279 @default.
• W2066231048 hasConcept C185592680 @default.
• W2066231048 hasConcept C2777622882 @default.
• W2066231048 hasConcept C2777742833 @default.
• W2066231048 hasConcept C2778326061 @default.
• W2066231048 hasConcept C2780870201 @default.
• W2066231048 hasConcept C29688787 @default.
• W2066231048 hasConcept C2992676626 @default.
• W2066231048 hasConcept C41895202 @default.
• W2066231048 hasConcept C519581460 @default.
• W2066231048 hasConcept C55493867 @default.
• W2066231048 hasConcept C86803240 @default.
• W2066231048 hasConcept C95444343 @default.
• W2066231048 hasConceptScore W2066231048C112243037 @default.
• W2066231048 hasConceptScore W2066231048C12554922 @default.
• W2066231048 hasConceptScore W2066231048C138885662 @default.
• W2066231048 hasConceptScore W2066231048C178790620 @default.
• W2066231048 hasConceptScore W2066231048C181199279 @default.
• W2066231048 hasConceptScore W2066231048C185592680 @default.
• W2066231048 hasConceptScore W2066231048C2777622882 @default.
• W2066231048 hasConceptScore W2066231048C2777742833 @default.
• W2066231048 hasConceptScore W2066231048C2778326061 @default.
• W2066231048 hasConceptScore W2066231048C2780870201 @default.
• W2066231048 hasConceptScore W2066231048C29688787 @default.
• W2066231048 hasConceptScore W2066231048C2992676626 @default.
• W2066231048 hasConceptScore W2066231048C41895202 @default.
• W2066231048 hasConceptScore W2066231048C519581460 @default.
• W2066231048 hasConceptScore W2066231048C55493867 @default.
• W2066231048 hasConceptScore W2066231048C86803240 @default.
• W2066231048 hasConceptScore W2066231048C95444343 @default.
• W2066231048 hasIssue "25" @default.
• W2066231048 hasLocation W20662310481 @default.
• W2066231048 hasOpenAccess W2066231048 @default.
• W2066231048 hasPrimaryLocation W20662310481 @default.
• W2066231048 hasRelatedWork W1980406827 @default.
• W2066231048 hasRelatedWork W1990796803 @default.
• W2066231048 hasRelatedWork W2055114314 @default.
• W2066231048 hasRelatedWork W2066231048 @default.
• W2066231048 hasRelatedWork W2094694252 @default.
• W2066231048 hasRelatedWork W2108271801 @default.
• W2066231048 hasRelatedWork W2243704036 @default.
• W2066231048 hasRelatedWork W2374639172 @default.
• W2066231048 hasRelatedWork W4313198982 @default.
• W2066231048 hasRelatedWork W2184702858 @default.

• next