CR Domains Research Articles

TEM Investigation of Asymmetric Deposition-Driven Crystalline-to-Amorphous Transition in Silicon Nanowires.

Controlling the shape and internal strain of nanowires (NWs) is critical for their safe and reliable use and for the exploration of novel functionalities of nanodevices. In this work, transmission electron microscopy was employed to examine bent Si NWs prepared by asymmetric electron-beam evaporation. The asymmetric deposition of Cr caused the formation of nanosized amorphous-Si domains; the non-crystallinity of the Si NWs was controlled by the bending radius. No other intermediate crystalline phase was present during the crystalline-to-amorphous transition, indicating a direct phase transition from the original crystalline phase to the amorphous phase. Moreover, amorphous microstructures caused by compressive stress, such as amorphous Cr domains and boxes, were also observed in the asymmetric Cr layer used to induce bending, and the local non-crystallinity of Cr was lower than that of Si under the same bending radius.

Enhanced strength-ductility synergy in a new 2.2 GPa grade ultra-high strength stainless steel with balanced fracture toughness: Elucidating the role of duplex aging treatment

Tuning thermal treatment schedule has been confirmed as an effective technical route for achieving optimum properties of steels. In the present work, the role of a unique duplex aging process on the enhanced strength-ductility synergy in a newly designed 2.2 GPa grade martensitic ultra-high strength stainless steel (UHSSS) was fully elucidated utilizing multi-scale characterization methods and various mechanical tests. Compared with the first aging-treated steel, the strength and ductility are synergistically enhanced after the second aging treatment due to increased volume fractions of Laves phase and reversed austenite as well as intensification of spinodal decomposition. Specifically, nanoprecipitates not only act as “obstacles” for dislocation movements, but also delay the deformation localization and necking instability, resulting in the dramatic increase of both strength (359 MPa for yield strength) and ductility (17% for reduction of area). The second aging-treated steel exhibited an impressive combination of mechanical properties: yield strength ∼ 1876 MPa, ultimate tensile strength ∼ 2259 MPa, elongation to fracture ∼ 11.5%, reduction of area ∼ 52%, and fracture toughness ∼ 43 MPa·m0.5 at ambient temperature. As revealed by the microstructural characterization, the aged steel is primarily strengthened by three distinct types of nanoprecipitates, including M2C, Laves phase, and α’Cr domains. The strengthening contributions of these precipitates were quantitatively estimated in terms of Orowan dislocation looping or cutting mechanisms. It is found that Laves phase contributed the maximum strength increment induced by precipitation hardening. An accessible pathway consisting of duplex aging treatment and co-precipitation of multiple nanoprecipitates is thus validated to exploit high-performance UHSSSs.

Structural Basis for the Interaction of Fibrin with the Very Low-Density Lipoprotein Receptor Revealed by NMR and Site-Directed Mutagenesis.

Interaction of fibrin with the very low-density lipoprotein receptor (VLDLR) promotes transendothelial migration of leukocytes and thereby inflammation. To establish the structural basis for this interaction, we have previously localized the VLDLR-binding site to fibrin βN-domains including fibrin β chain sequence 15-64 and determined the NMR solution structure of the VLDLR(2-4) fragment containing fibrin-binding CR domains 2-4 of VLDLR. In this study, we identified amino acid residues in VLDLR and the βN-domains that are involved in the interaction using NMR and site-directed mutagenesis. The results obtained revealed that Lys47 and Lys53 of the second and third positively charged clusters of the βN-domain, respectively, interact with Trp20 and Asp25 of the CR2 domain and Trp63 and Glu68 of the CR3 domain, respectively. This finding indicates that Lys residues of the βN-domain interact with the Lys-binding site of the CR domains in a manner proposed earlier for the interaction of other members of the LDL receptor family with their ligands. In addition, Gly15 of the βN-domain and its first positively charged cluster contribute to the high-affinity interaction with VLDLR. Molecular modeling based on the results obtained and analysis of the previously published structures of such domains complexed with RAP and HRV2 allowed us to propose a model of interaction of fibrin βN-domains with the fibrin-binding CR domains of the VLDL receptor.

Analysis of E1A domains involved in the enhancement of CDK2 activity

E1A is an adenoviral protein which is expressed at the early phase after viral infection and contains four conserved regions (CR1, CR2, CR3 and CR4). Our previous work suggests that E1A facilitates the formation of cyclin A-CDK2 complex and thereby enhances CDK2 activity. However, the molecular function of E1A in CDK2 activation has been unclear. Here, we studied the mechanism of enhancement of CDK2 activity by E1A, using the E1A variant forms which selectively contain CR domains. We isolated four E1A variant forms, i.e. 13S (containing CR1, CR2, CR3, CR4), 12S (CR1, CR2, CR4), 10S (CR2, CR4) and 9S (CR4), derived from HEK293 cells which express E1A. 13S promoted G2/M-phase arrest, upon CDK2 hyper-activation by co-expressing a stabilized cyclin A mutant, most strongly among those E1A variant forms. Concomitantly, the specific activity of the 13S-associated CDK2 was highest among them. 10S exhibited lower affinity for CDK2 than the 13S while the affinity for CDK2 was comparable between 13S and 12S. Nonetheless, 12S did not enhance the CDK2 specific activity. On the other hand, a mutation in CR2 domain, which is essential for binding to p107, suppressed both the binding and activation of CDK2. These results suggest that CR1 domain, in addition to CR2 domain via p107 interaction, is important for binding to CycA-CDK2 complex while CR3 domain facilitates CDK2 activation. Since the function of CR3 in cell cycle regulation has been relatively unknown, we propose the enhancement of CDK2 activity as a novel function of CR3 domain.

Structural basis of the activation of type 1 insulin-like growth factor receptor

Type 1 insulin-like growth factor receptor (IGF1R) is a receptor tyrosine kinase that regulates cell growth and proliferation, and can be activated by IGF1, IGF2, and insulin. Here, we report the cryo-EM structure of full-length IGF1R–IGF1 complex in the active state. This structure reveals that only one IGF1 molecule binds the Γ-shaped asymmetric IGF1R dimer. The IGF1-binding site is formed by the L1 and CR domains of one IGF1R protomer and the α-CT and FnIII-1 domains of the other. The liganded α-CT forms a rigid beam-like structure with the unliganded α-CT, which hinders the conformational change of the unliganded α-CT required for binding of a second IGF1 molecule. We further identify an L1–FnIII-2 interaction that mediates the dimerization of membrane-proximal domains of IGF1R. This interaction is required for optimal receptor activation. Our study identifies a source of the negative cooperativity in IGF1 binding to IGF1R and reveals the structural basis of IGF1R activation.

Binding and Synergizing Motif within Coleopteran Cadherin Enhances Cry3Bb Toxicity on the Colorado Potato Beetle and the Lesser Mealworm.

Cry3Bb toxin from Bacillus thuringiensis is an important insecticidal protein due to its potency against coleopteran pests, especially rootworms. Cadherin, a protein in the insect midgut epithelium, is a receptor of Cry toxins; in some insect species toxin-binding domains of cadherins-synergized Cry toxicity. Previously, we reported that the DvCad1-CR8-10 fragment of Diabrotica virgifera virgifera cadherin-like protein (GenBank Accession #EF531715) enhanced Cry3Bb toxicity to the Colorado Potato Beetle (CPB), Leptinotarsa decimlineata (L. decimlineata). We report that individual CR domains of the DvCad1-CR8-10 fragment were found to have strong binding affinities to α-chymotrypsin-treated Cry3Bb. The dissociation constant (Kd) of Cry3Bb binding to the CR8, CR9, and CR10 domain was 4.9 nM, 28.2 nM, and 4.6 nM, respectively. CR8 and CR10, but not CR9, enhanced Cry3Bb toxicity against L. decimlineata and the lesser mealworm Alphitobius diaperinus neonates. In-frame deletions of the DvCad1-CR10 open reading frame defined a high-affinity binding and synergistic site to a motif in residues I1226–D1278. A 26 amino acid peptide from the high affinity Cry3Bb-binding region of CR10 functioned as a Cry3Bb synergist against coleopteran larvae.

Structural basis for the heme transfer reaction in heme uptake machinery from Corynebacteria.

The crystal structures of the conserved region domains of HtaA and HtaB, which act as heme binding/transport proteins in the heme uptake machinery in Corynebacterium glutamicum, are determined for the first time. The molecular mechanism of heme transfer among these proteins is proposed based on the spectroscopic and structural analyses.

Structural basis for the recognition of LDL-receptor family members by VSV glycoprotein

Vesicular stomatitis virus (VSV) is an oncolytic rhabdovirus and its glycoprotein G is widely used to pseudotype other viruses for gene therapy. Low-density lipoprotein receptor (LDL-R) serves as a major entry receptor for VSV. Here we report two crystal structures of VSV G in complex with two distinct cysteine-rich domains (CR2 and CR3) of LDL-R, showing that their binding sites on G are identical. We identify two basic residues on G, which are essential for its interaction with CR2 and CR3. Mutating these residues abolishes VSV infectivity even though VSV can use alternative receptors, indicating that all VSV receptors are members of the LDL-R family. Collectively, our data suggest that VSV G has specifically evolved to interact with receptor CR domains. These structural insights into the interaction between VSV G and host cell receptors provide a basis for the design of recombinant viruses with an altered tropism.

Interaction of Fibrin with the Very Low-Density Lipoprotein (VLDL) Receptor: Further Characterization and Localization of the VLDL Receptor-Binding Site in Fibrin βN-Domains.

Our recent study revealed that fibrin and the very low-density lipoprotein receptor (VLDLR) interact with each other through a pair of fibrin βN-domains and CR domains of the receptor and this interaction promotes transendothelial migration of leukocytes and thereby inflammation. The major objectives of this study were to further clarify the molecular mechanism of fibrin-VLDLR interaction and to identify amino acid residues in the βN-domains involved in this interaction. Our binding experiments with the (β15-66)2 fragment, which corresponds to a pair of fibrin βN-domains, and the VLDLR(1-8) fragment, consisting of eight CR domains of VLDLR, revealed that interaction between them strongly depends on ionic strength and chemical modification of all Lys or Arg residues in (β15-66)2 results in abrogation of this interaction. To identify which of these residues are involved in the interaction, we mutated all Lys or Arg residues in each of the three positively charged Lys/Arg clusters of the (β15-66)2 fragment, as well as single Arg17 and Arg30, and tested the affinity of the mutants obtained for VLDLR(1-8) by an enzyme-linked immunosorbent assay and surface plasmon resonance. The experiments revealed that the second and third Lys/Arg clusters make the major contribution to this interaction while the contribution of the first cluster is moderate. The results obtained suggest that interaction between fibrin and the VLDL receptor employs the "double-Lys/Arg" recognition mode previously proposed for the interaction of the LDL receptor family members with their ligands. They also provide valuable information for the development of highly specific peptide-based inhibitors of fibrin-VLDLR interaction.

ADAM and ADAMTS Family Proteins and Snake Venom Metalloproteinases: A Structural Overview.

A disintegrin and metalloproteinase (ADAM) family proteins constitute a major class of membrane-anchored multidomain proteinases that are responsible for the shedding of cell-surface protein ectodomains, including the latent forms of growth factors, cytokines, receptors and other molecules. Snake venom metalloproteinases (SVMPs) are major components in most viper venoms. SVMPs are primarily responsible for hemorrhagic activity and may also interfere with the hemostatic system in envenomed animals. SVMPs are phylogenetically most closely related to ADAMs and, together with ADAMs and related ADAM with thrombospondin motifs (ADAMTS) family proteinases, constitute adamalysins/reprolysins or the M12B clan (MEROPS database) of metalloproteinases. Although the catalytic domain structure is topologically similar to that of other metalloproteinases such as matrix metalloproteinases, the M12B proteinases have a modular structure with multiple non-catalytic ancillary domains that are not found in other proteinases. Notably, crystallographic studies revealed that, in addition to the conserved metalloproteinase domain, M12B members share a hallmark cysteine-rich domain designated as the “ADAM_CR” domain. Despite their name, ADAMTSs lack disintegrin-like structures and instead comprise two ADAM_CR domains. This review highlights the current state of our knowledge on the three-dimensional structures of M12B proteinases, focusing on their unique domains that may collaboratively participate in directing these proteinases to specific substrates.

Interaction of Fibrin with the Very Low Density Lipoprotein Receptor: Further Characterization and Localization of the Fibrin-Binding Site.

Our recent study revealed that fibrin interacts with the very low density lipoprotein receptor (VLDLR) on endothelial cells through its βN domains, and this interaction promotes transendothelial migration of leukocytes and thereby inflammation. The major aims of this study were to further characterize this interaction and localize the fibrin-binding site in the VLDLR. To localize the fibrin-binding site, we expressed a soluble extracellular portion of this receptor, sVLDLRHT, its N- and C-terminal regions, VLDLR(1-8)HT and des(1-8)VLDLRHT, respectively, and a number of VLDLR fragments containing various combinations of CR domains and confirmed their proper folding by fluorescence spectroscopy. Interaction of these fragments with the (β15-66)2 fragment corresponding to a pair of VLDLR-binding βN domains of fibrin was tested by different methods. Our experiments performed by an enzyme-linked immunosorbent assay and surface plasmon resonance revealed that the VLDLR(1-8)HT fragment containing eight CR domains of VLDLR and its subfragments, VLDLR(1-4)HT and VLDLR(2-4)HT, interact with (β15-66)2 with practically the same affinity as sVLDLRHT while the affinity of VLDLR(2-3)HT was ∼2-fold lower. In contrast, des(1-8)VLDLRHT exhibited no binding. Formation of the complex in solution between the fibrin-binding fragments of VLDLR and (β15-66)2 was detected by fluorescence spectroscopy. In addition, formation of a complex between VLDLR(2-4)HT and (β15-66)2 in solution was confirmed by size-exclusion chromatography. Thus, the results obtained indicate that minimal fibrin-binding structures are located within the second and third CR domains of the VLDL receptor and the presence of the fourth CR domain is required for high-affinity binding. They also indicate that tryptophan residues of CR domains are involved in this binding.

Growth of antiferromagnetically ordered Cr monolayer on Ag(100)

Two-dimensional antiferromagnetism observed on flat Cr monolayers deposited on Ag(100) has been investigated under various growth conditions. Cr monolayer domains are found to grow within the temperature range of 373–453 K while multilayer growth mode is prevalent at room temperature (RT). In agreement with theoretical predictions, a c(2 × 2) antiferromagnetic configuration is observed on Cr monolayer and is confirmed by the presence of magnetic superstructural spots in Low Energy Electron Diffraction (LEED) and antiferromagnetic Cr 3d bands in Angle-resolved Photoemission Spectroscopy (ARPES) studies. Antiferromagnetism of Cr monolayer film is found to depend strongly on various growth parameters such as growth temperature, annealing temperature, annealing duration, rate of growth etc. and are studied in detail here. The phenomenon of Cr–Ag intermixing at the interface is found to be the main factor for the complex growth of Cr on Ag(100) substrate. Optimum parameters for the growth of large monolayer Cr domains on Ag(100) substrate are presented here.

Quantitative Dissection of the Binding Contributions of Ligand Lysines of the Receptor-associated Protein (RAP) to the Low Density Lipoprotein Receptor-related Protein (LRP1)

Although lysines are known to be critical for ligand binding to LDL receptor family receptors, relatively small reductions in affinity have been found when such lysines have been mutated. To resolve this paradox, we have examined the specific binding contributions of four lysines, Lys-253, Lys-256, Lys-270, and Lys-289, in the third domain (D3) of receptor-associated protein (RAP), by eliminating all other lysine residues. Using D3 variants containing lysine subsets, we examined binding to the high affinity fragment CR56 from LRP1. With this simplification, we found that elimination of the lysine pairs Lys-253/Lys-256 and Lys-270/Lys-289 resulted in increases in Kd of 1240- and 100,000-fold, respectively. Each pair contributed additively to overall affinity, with 61% from Lys-270/Lys-289 and 39% from Lys-253/Lys-256. Furthermore, the Lys-270/Lys-289 pair alone could bind different single CR domains with similar affinity. Within the pairs, binding contributions of Lys-270 ≫ Lys-256 > Lys-253 ∼ Lys-289 were deduced. Importantly, however, Lys-289 could significantly compensate for the loss of Lys-270, thus explaining how previous studies have underestimated the importance of Lys-270. Calorimetry showed that favorable enthalpy, from Lys-256 and Lys-270, overwhelmingly drives binding, offset by unfavorable entropy. Our findings support a mode of ligand binding in which a proximal pair of lysines engages the negatively charged pocket of a CR domain, with two such pairs of interactions (requiring two CR domains), appropriately separated, being alone sufficient to provide the low nanomolar affinity found for most protein ligands of LDL receptor family members.

Role of Dynamics in the Autoinhibition and Activation of the Exchange Protein Directly Activated by Cyclic AMP (EPAC)

The exchange protein directly activated by cAMP (EPAC) is a key receptor of cAMP in eukaryotes and controls critical signaling pathways. Currently, no residue resolution information is available on the full-length EPAC dynamics, which are known to be pivotal determinants of allostery. In addition, no information is presently available on the intermediates for the classical induced fit and conformational selection activation pathways. Here these questions are addressed through molecular dynamics simulations on five key states along the thermodynamic cycle for the cAMP-dependent activation of a fully functional construct of EPAC2, which includes the cAMP-binding domain and the integral catalytic region. The simulations are not only validated by the agreement with the experimental trends in cAMP-binding domain dynamics determined by NMR, but they also reveal unanticipated dynamic attributes, rationalizing previously unexplained aspects of EPAC activation and autoinhibition. Specifically, the simulations show that cAMP binding causes an extensive perturbation of dynamics in the distal catalytic region, assisting the recognition of the Rap1b substrate. In addition, analysis of the activation intermediates points to a possible hybrid mechanism of EPAC allostery incorporating elements of both the induced fit and conformational selection models. In this mechanism an entropy compensation strategy results in a low free-energy pathway of activation. Furthermore, the simulations indicate that the autoinhibitory interactions of EPAC are more dynamic than previously anticipated, leading to a revised model of autoinhibition in which dynamics fine tune the stability of the autoinhibited state, optimally sensitizing it to cAMP while avoiding constitutive activation.

Dissection of RAP-LRP interactions: Binding of RAP and RAP fragments to complement-like repeats 7 and 8 from ligand binding cluster II of LRP

The receptor associated protein (RAP) is a three domain 38 kDa ER-resident chaperone that helps folding of LRP and other LDL receptor family members and prevents premature binding of protein ligands. It competes strongly with all known LRP ligands. To further understanding of the specificity of RAP-LRP interactions, the binding of RAP and RAP fragments to two domains (CR7–CR8) from one of the main ligand-binding regions of LRP has been examined by 2D HSQC NMR spectroscopy and isothermal titration calorimetry. We found that RAP contains two binding sites for CR7–CR8, with the higher affinity site ( K d ∼ 1 μM) located in the C-terminal two-thirds and the weaker site ( K d ∼ 5 μM) in the N-terminal third of RAP. Residues from both CR7 and CR8 are involved in binding at each RAP site. The presence of more than one binding site on RAP for CR domains from LRP, together with the previous demonstration by others that RAP can bind to CR5–CR6 with comparably low affinities suggest an explanation for the dual roles of RAP as a folding chaperone and a tight competitive inhibitor of ligand binding.

Surface analytical chemical imaging and morphology of Cu–Cr alloy

Cu–Cr alloys are very interesting materials to study non-equilibrium phase transformations in binary systems containing immiscible metals as well as for practical applications as high-thermal conductivity and contact materials. We studied by high resolution secondary ion mass spectrometry and atomic force microscopy the chemical element distributions and surface morphology of Cu–Cr alloys irradiated with a low energy, high current electron beam. The irradiation conditions were: energy density 4 ÷ 6 J cm − 2, pulse duration 2.8 ÷ 3.6 μs, number of shots 16. Cu–Cr alloys were prepared by powder metallurgy and contain 30 wt.% Cr. While in samples irradiated at low energy density maps of surface elements reveal the presence of well separated micrometer-sized domains of Cr and Cu, after irradiation at high energy density sub-micron spots of coexisting Cu and Cr appear. This is a clear indication of CuCr compound formation, as also supported by local mass spectrometry. Irradiated samples have reduced roughness, especially in Cr domains, with respect to the pre-treated sample. However, microcracks and microcraters appear as beam energy density and number of shots increase, thus compromising the smoothening effect and the surface integrity.

Altering the Alignment of the Active Site of Factor VIIa by Raising Tissue Factor above the Phospholipid Surface.

Tissue factor (TF), an integral membrane protein, is the protein cofactor for coagulation factor VIIa (VIIa). The membrane-bound enzyme complex of TF and VIIa (TF:VIIa) initiates coagulation by activating factor X to Xa via limited proteolysis. Free VIIa has limited enzymatic activity, but when in complex with TF, its activity -- particularly its ability to cleave macromolecular substrates -- dramatically increases. VIIa in complex with truncated, soluble TF (sTF; consisting of the isolated extracellular domain) has much lower procoagulant activity than VIIa in complex with membrane-anchored TF in a suitable phospholipid membrane. When TF binds to VIIa, it re-aligns the active site of VIIa relative to the membrane surface, and it is hypothesized that one of the essential functions of protein cofactors in the clotting cascade is to position the active sites of their cognate proteases the proper distance and orientation above the membrane surface for optimal attack on the scissile bonds of membrane-bound protein substrates. We are testing how this alignment contributes to catalysis by altering the distance of TF above the phospholipid surface, thereby altering the position of VIIa's active site above the membrane. We created a number of chimeras between the extracellular domain of TF and portions of the integral membrane protein, P-selectin, in order to raise TF varying distances above the phospholipid surface. TFCR9 is the shortest chimera, containing the 9th consensus repeat domain (CR) and transmembrane domain of P-selectin. TFCR8.9 contains the 8th and 9th CR domains as well as the transmembrane domain of P-selectin. TFLCR8.9 is identical to TFCR8.9 except that a flexible linker of four glycines and a serine residue repeated three times, (G4S)3, separates the TF and P-selectin sequences. All of the chimeras allosterically activated VIIa to the same extent that sTF or TF did (measured by cleavage of small peptidyl-amide substrates) confirming that the chimeras are properly folded. VIIa bound to the chimeras with Kd values ranging from 2.6 to 4.2 nM, which is equivalent to its interaction with sTF (Kd, 2-5 nM) rather than with TF in phospholipid membranes (Kd < 50 pM). This also confirms that the chimeras are properly folded and strongly suggests that binding interactions do not take place between VIIa and the membrane surface when VIIa binds to these chimeras. The chimera:VIIa complexes had reduced ability to activate X compared to TF:VIIa complexes, and the reduction was greater with the longer chimera, TFCR8.9, demonstrating a three-fold reduction in activity compared to TF. The chimeras had greatly reduced procoagulant activity compared to TF, with only 6-10% the specific activity of TF. Interestingly, the chimeras had significantly higher procoagulant activity if the chimera:VIIa complexes were pre-assembled before adding plasma, suggesting that the chimeras may have a defect in capturing VII and/or supporting its conversion to VIIa. These results imply that maintaining the proper distance between TF and the membrane surface is important not only for TF:VIIa to activate macromolecular substrates, but possibly also for TF to capture and bind membrane-bound VII and/or VIIa.

Photoemission and LEED investigation of the initial growth of Cr on Ag( [formula omitted

The initial growth of Cr on a Ag(0 0 1) surface is studied by low energy electron diffraction (LEED), angle resolved ultra-violet photoemission and X-ray photoelectron diffraction, as a function of deposition temperature. At 100 K, the LEED diagram is p(1×1) and a highly defected monolayer is initially formed. When the substrate is kept at room temperature during Cr evaporation, the LEED pattern is still p(1×1), and multilayer growth with markedly improved atomic order and accompanied by Ag segregation occurs. In contrast, at 430 K, a weak c(2×2) LEED diagram is obtained at low electron energies (<85 eV), attributed to the presence of well ordered bare Cr monolayer islands with p(1×1) atomic order but presenting a c(2×2) antiferromagnetic superstructure. This superstructure is identical in every respect to the one identified in our earlier work [Phys. Rev. B 44 (1991) 8407] and quite similar to the magnetic c(2×2) Mn monolayer on Ag(0 0 1) investigated recently by dynamical LEED [Phys. Rev. B 62 (2000) 2944]. Valence band data indicate coexistence of monolayer c(2×2) Cr domains and multilayer patches made of p(1×1) Ag islands on top of a mixed Cr–Ag phase.

BMP-binding modules in chordin: a model for signalling regulation in the extracellular space.

A number of genetic and molecular studies have implicated Chordin in the regulation of dorsoventral patterning during gastrulation. Chordin, a BMP antagonist of 120 kDa, contains four small (about 70 amino acids each) cysteine-rich domains (CRs) of unknown function. In this study, we show that the Chordin CRs define a novel protein module for the binding and regulation of BMPs. The biological activity of Chordin resides in the CRs, especially in CR1 and CR3, which have dorsalizing activity in Xenopus embryo assays and bind BMP4 with dissociation constants in the nanomolar range. The activity of individual CRs, however, is 5- to 10-fold lower than that of full-length Chordin. These results shed light on the molecular mechanism by which Chordin/BMP complexes are regulated by the metalloprotease Xolloid, which cleaves in the vicinity of CR1 and CR3 and would release CR/BMP complexes with lower anti-BMP activity than intact Chordin. CR domains are found in other extracellular proteins such as procollagens. Full-length Xenopus procollagen IIA mRNA has dorsalizing activity in embryo microinjection assays and the CR domain is required for this activity. Similarly, a C. elegans cDNA containing five CR domains induces secondary axes in injected Xenopus embryos. These results suggest that CR modules may function in a number of extracellular proteins to regulate growth factor signalling.

Crystallographic and electronic structure of Cu/Cr/Cu(001) sandwiches

Thin Cu layers have been evaporated at room temperature on Cr films (~ 10 monolayers) deposited on a Cu(001) single crystal. The Crystallographic structure of these sandwiches is analysed by means of low-energy electron diffraction (LEED), angle-resolved ultra-violet photoemission spectroscopy (ARUPS) and X-ray photoemission spectroscopy. The data show that the attenuation of the Cr signal versus Cu deposition is much lower than expected for a uniform film growth. Moreover, the LEED pattern characteristic of the Cr film (made of Cr(110) domains with four different orientations) is visible for Cu coverages as high as 23 ML. This indicates a strong diffusion of the Cu atoms on the Cr film surface and the formation of Cu three-dimensional islands which present, according to ARUPS and X-ray photoelectron diffraction measurements, a highly disordered fcc Crystallographic structure. However the low reactivity of the sandwiches towards oxygen suggests that the whole Cr surface is wetted by at least 1 ML of Cu.