Neutron scattering study of the (γ-B) catalytic domains of complement proteases Cl̄r and Cl̄s

AbstractThe catalytic domains of Cl̄r and Cl̄s, comprising the C-tenninal region of the A chain (γ), disulphide-linked to the B chain, were obtained by limited proteolysis of the native proteases with chymotrypsin and plasmin, respectively, and studied by small angle neutron scattering. For CIs (γ-B), a molar mass of 45 000 ± 5000 gmol, and a relatively large radius of gyration (Rg) of 28 ± 1 Å were determined, excluding a single globular domain. The corresponding values for Cl̄r (γ-B), (90,000 gmol, Rg, = 34 ± 1 Å) are consistent with a dimer involving the loose packing of two (γ-B) subunits. Various models of the dimer are discussed in the light of neutron scattering and other data

<Session 3: Biomedical Technology>Assessing radio frequency electromagnetic field exposure with a wearable network of dosimeters

19–22 May 2022 Kyoto, JapanIn contrast to the tremendous increase of wireless applications, the knowledge about daily life radio frequency electromagnetic field (RF-EMF) exposure remains low. Some research has already been conducted using large commercial single-antenna on-body dosimeters to assess daily RF-EMF exposure in different environments. To increase measurement quality and decrease variability however, a distributed, wearable body sensor network spread over the body for the assessment of RF-EMF exposure is desired. As a better alternative to the common single node assessment technique, this work therefore presents a wearable sensor network consisting of five nodes for the assessment of exposure in the 389 to 464, 779 to 950 and 2400 to 2483.5MHz bands using only two transceivers per node. A single node supports an antenna connection for the first band, two for the second and two for the third band. This makes antenna diversity possible and hence increases sensitivity for specific frequencies, depending on the choice of antenna design. Every node is powered by two AAA batteries, which define the size of the system (53x25x15mm), making it smaller than any other commercially available dosimeter. Furthermore, the device supports an inertial sensor for the assessment of body posture and/or activity during the measurement

Channel gain for a wrist-to-arm scenario in the 55-65 GHz frequency band

Wireless communication on the body is expected to become more important in the future. This communication will in certain scenarios benefit from higher frequencies of operation and their associated smaller antennas and potentially higher bandwidths. One of these scenarios is communication between a wristband and wearable sensors on the arm. In order to investigate the feasibility of such a scenario, propagation at 55â65 GHz along the arm is measured for two configurations. First, for increasing separation distances along the arm, and second for a transmitter is rotationally placed around the wrist. Two channel gain models are fitted to the data and used to obtain a channel gain exponent in the first configuration and loss per angle of rotation in the second configuration. These models are relevant inputs for the design of future wearable wireless systems

Cosmic rays and the primordial gas

One of the most outstanding problems in the gravitational collapse scenario of early structure formation is the cooling of primordial gas to allow for small mass objects to form. As the neutral primordial gas is a poor radiator at temperatures (T\le10^4\unit{K}), molecular hydrogen is needed for further cooling down to temperatures (T\sim100\unit{K}). The formation of molecular hydrogen is catalyzed by the presence of free electrons, which could be provided by the ionization due to an early population of cosmic rays. In order to investigate this possibility we developed a code to study the effects of ionizing cosmic rays on the thermal and chemical evolution of primordial gas. We found that cosmic rays can provide enough free electrons needed for the formation of molecular hydrogen, and therefore can increase the cooling ability of such primordial gas under following conditions: A dissociating photon flux with (F<10^{-18}\unit{erg cm^{-2} Hz^{-1} s^{-1}}), initial temperature of the gas (\sim10^{3}\unit{K}), total gas number densities (n\ge1\unit{cm^{-3}}), Cosmic ray sources with (\dot{\epsilon}_{CR}>10^{-33}\unit{erg cm^{-3} s^{-1}}).Comment: 15 pages, 5 figure

The serine protease domain of MASP-3: enzymatic properties and crystal structure in complex with ecotin.

International audienceMannan-binding lectin (MBL), ficolins and collectin-11 are known to associate with three homologous modular proteases, the MBL-Associated Serine Proteases (MASPs). The crystal structures of the catalytic domains of MASP-1 and MASP-2 have been solved, but the structure of the corresponding domain of MASP-3 remains unknown. A link between mutations in the MASP1/3 gene and the rare autosomal recessive 3MC (Mingarelli, Malpuech, Michels and Carnevale,) syndrome, characterized by various developmental disorders, was discovered recently, revealing an unexpected important role of MASP-3 in early developmental processes. To gain a first insight into the enzymatic and structural properties of MASP-3, a recombinant form of its serine protease (SP) domain was produced and characterized. The amidolytic activity of this domain on fluorescent peptidyl-aminomethylcoumarin substrates was shown to be considerably lower than that of other members of the C1r/C1s/MASP family. The E. coli protease inhibitor ecotin bound to the SP domains of MASP-3 and MASP-2, whereas no significant interaction was detected with MASP-1, C1r and C1s. A tetrameric complex comprising an ecotin dimer and two MASP-3 SP domains was isolated and its crystal structure was solved and refined to 3.2 Å. Analysis of the ecotin/MASP-3 interfaces allows a better understanding of the differential reactivity of the C1r/C1s/MASP protease family members towards ecotin, and comparison of the MASP-3 SP domain structure with those of other trypsin-like proteases yields novel hypotheses accounting for its zymogen-like properties in vitro

Structure of the C1r-C1s interaction of the C1 complex of complement activation.

The multiprotein complex C1 initiates the classical pathway of complement activation on binding to antibody-antigen complexes, pathogen surfaces, apoptotic cells, and polyanionic structures. It is formed from the recognition subcomponent C1q and a tetramer of proteases C1r2C1s2 as a Ca2+-dependent complex. Here we have determined the structure of a complex between the CUB1-EGF-CUB2 fragments of C1r and C1s to reveal the C1r-C1s interaction that forms the core of C1. Both fragments are L-shaped and interlock to form a compact antiparallel heterodimer with a Ca2+ from each subcomponent at the interface. Contacts, involving all three domains of each protease, are more extensive than those of C1r or C1s homodimers, explaining why heterocomplexes form preferentially. The available structural and biophysical data support a model of C1r2C1s2 in which two C1r-C1s dimers are linked via the catalytic domains of C1r. They are incompatible with a recent model in which the N-terminal domains of C1r and C1s form a fixed tetramer. On binding to C1q, the proteases become more compact, with the C1r-C1s dimers at the center and the six collagenous stems of C1q arranged around the perimeter. Activation is likely driven by separation of the C1r-C1s dimer pairs when C1q binds to a surface. Considerable flexibility in C1s likely facilitates C1 complex formation, activation of C1s by C1r, and binding and activation of downstream substrates C4 and C4b-bound C2 to initiate the reaction cascade.Funding for this work was provided by the Medical Research Council (Grant G1000191/1, to R.W., P.C.E.M., and W.J.S.)

Periodontal Ehlers-Danlos Syndrome Is Caused by Mutations in C1R and C1S, which Encode Subcomponents C1r and C1s of Complement

Periodontal Ehlers-Danlos syndrome (pEDS) is an autosomal-dominant disorder characterized by early-onset periodontitis leading to premature loss of teeth, joint hypermobility, and mild skin findings. A locus was mapped to an approximately 5.8 Mb region at 12p13.1 but no candidate gene was identified. In an international consortium we recruited 19 independent families comprising 107 individuals with pEDS to identify the locus, characterize the clinical details in those with defined genetic causes, and try to understand the physiological basis of the condition. In 17 of these families, we identified heterozygous missense or in-frame insertion/deletion mutations in C1R (15 families) or C1S (2 families), contiguous genes in the mapped locus that encode subunits C1r and C1s of the first component of the classical complement pathway. These two proteins form a heterotetramer that then combines with six C1q subunits. Pathogenic variants involve the subunit interfaces or inter-domain hinges of C1r and C1s and are associated with intracellular retention and mild endoplasmic reticulum enlargement. Clinical features of affected individuals in these families include rapidly progressing periodontitis with onset in the teens or childhood, a previously unrecognized lack of attached gingiva, pretibial hyperpigmentation, skin and vascular fragility, easy bruising, and variable musculoskeletal symptoms. Our findings open a connection between the inflammatory classical complement pathway and connective tissue homeostasis

Borrelia burgdorferi BBK32 Inhibits the Classical Pathway by Blocking Activation of the C1 Complement Complex

Citation: Garcia, B. L., Zhi, H., Wager, B., Hook, M., & Skare, J. T. (2016). Borrelia burgdorferi BBK32 Inhibits the Classical Pathway by Blocking Activation of the C1 Complement Complex. Plos Pathogens, 12(1), 28. doi:10.1371/journal.ppat.1005404Pathogens that traffic in blood, lymphatics, or interstitial fluids must adopt strategies to evade innate immune defenses, notably the complement system. Through recruitment of host regulators of complement to their surface, many pathogens are able to escape complement-mediated attack. The Lyme disease spirochete, Borrelia burgdorferi, produces a number of surface proteins that bind to factor H related molecules, which function as the dominant negative regulator of the alternative pathway of complement. Relatively less is known about how B. burgdorferi evades the classical pathway of complement despite the observation that some sensu lato strains are sensitive to classical pathway activation. Here we report that the borrelial lipoprotein BBK32 potently and specifically inhibits the classical pathway by binding with high affinity to the initiating C1 complex of complement. In addition, B. burgdorferi cells that produce BBK32 on their surface bind to both C1 and C1r and a serum sensitive derivative of B. burgdorferi is protected from killing via the classical pathway in a BBK32-dependent manner. Subsequent biochemical and biophysical approaches localized the anti-complement activity of BBK32 to its globular C-terminal domain. Mechanistic studies reveal that BBK32 acts by entrapping C1 in its zymogen form by binding and inhibiting the C1 subcomponent, C1r, which serves as the initiating serine protease of the classical pathway. To our knowledge this is the first report of a spirochetal protein acting as a direct inhibitor of the classical pathway and is the only example of a biomolecule capable of specifically and noncovalently inhibiting C1/C1r. By identifying a unique mode of complement evasion this study greatly enhances our understanding of how pathogens subvert and potentially manipulate host innate immune systems