Intervertebral disc disease is one of the most common musculoskeletal disorders. A number environmental and anthropometric risk factors may contribute to it, recent reports have suggested importance genetic as well. The COL9A2 gene, which codes for polypeptide chains collagen IX that expressed in intervertebral disc, was screened sequence variations individuals with disease. analysis identified a putative disease-causing variation converted codon glutamine tryptophan six out 157 but none 174...

Previously, an assay called conformation sensitive gel electrophoresis (CSGE) was developed for scanning PCR products the presence of single-base and larger base mismatches in DNA. The based on assumption that mildly denaturing solvents appropriate buffer can accentuate conformational changes produced by double-stranded DNA thereby increase differential migration electrophoretic gels heteroduplexes homoduplexes. Here sensitivity assays CSGE improved limiting maximal size to 450 bp making...

Mutations in type 3 repeats of cartilage oligomeric matrix protein (COMP) cause two skeletal dysplasias, pseudoachondroplasia (PSACH) and multiple epiphyseal dysplasia (MED). We expressed recombinant wild-type COMP that showed structural functional properties identical to isolated from cartilage. A fragment encompassing the eight binds 14 calcium ions with moderate affinity high cooperativity presumably forms one large disulfide-bonded folding unit. PSACH mutant which Asp-469 was deleted...

The original patient with the Weissenbacher-Zweymüller syndrome was analyzed for mutations in two candidate genes expressed cartilage (COL2A1 and COL11A2). No were found COL2A1 gene but COL11A2 contained a single-base mutation that converted codon an obligate glycine to glutamate at position α2-955 (G955E). results here those published previously indicate (heterozygous OSMED), nonocular Stickler syndrome, homozygous OSMED are all caused by gene. Am. J. Med. Genet. 80:115–120, 1998. © 1998...

During atherogenesis, the extracellular pH of atherosclerotic lesions decreases. Here, we examined effect low, but physiologically plausible on aggregation modified LDL, one key processes in atherogenesis. LDL was treated with SMase, and SMase-treated followed at 5.5-7.5. The lower pH, more extensive identically prelipolyzed particles. At 5.5-6.0, aggregates were much larger (size >1 µm) than those formed neutral (100-200 nm). SMase treatment found to lead a dramatic decrease α-helix...

The N-terminal NC4 domain of collagen IX is a globular structure projecting away from the surface cartilage fibril. Several interactions have been suggested for this domain, reflecting its location and characteristic high isoelectric point. In an attempt to characterize in more detail, we set up prokaryotic expression system produce domain. purified 27.5-kDa product was analyzed glycosaminoglycan-binding potential by plasmon resonance solid-state assays. results show that specifically binds...

Thrombospondin-related anonymous protein, TRAP, has a critical role in the hepatocyte invasion step of Plasmodium sporozoites, transmissible form parasite causing malaria. The extracellular domains this sporozoite surface protein interact with receptors whereas its intracellular domain acts as link to actomyosin motor system. Liver heparan sulfate proteoglycans have been identified potential ligands for TRAP. Proteoglycan binding associated A- and TSR We present solution NMR structure TRAP...

The human COL11A2 gene was analyzed from two overlapping cosmid clones that were previously isolated in the course of searching major histocompatibility region (Janatipour, M., Naumov, Y., Ando, A., Sugimura, K., Okamoto, N., Tsuji, Abe, and Inoko, H.(1992) Immunogenetics 35, 272-278). Nucleotide sequencing defined over 28,000 base pairs gene. It shown to contain 66 exons. As with most genes for fibrillar collagens, first intron among largest, introns at 5′-end general larger than 3′-end....

HDL particles may enter atherosclerotic lesions having an acidic intimal fluid. Therefore, we investigated whether pH would affect their structural and functional properties. For this purpose, HDL2 HDL3 subfractions were incubated for various periods of time at different values ranging from 5.5 to 7.5, after which protein lipid compositions, size, structure, cholesterol efflux capacity analyzed. Incubation either subfraction induced unfolding apolipoproteins, was followed by release...

One of the major responses upon orexin receptor activation is Ca(2+) influx, and this influx seems to amplify other mediated by receptors. However, reduction in , often used assess importance might affect properties, like ligand-receptor interactions, as suggested for some GPCR systems. Hence, we investigated role interaction signal cascades apparent requirement orexin-A signalling.

As type IX collagen is a minor cartilage component, it difficult to purify sufficient amounts of from tissues or cultured cells study its structure and function. Also, the conventional pepsin digestion used for fibrillar collagens cannot be utilized purifying collagen, because contains several interruptions in collagenous triple helix. A baculovirus expression system was here produce recombinant human by coinfecting insect with three viruses containing full-length cDNAs α1(IX), α2(IX),...

Objective— In the deep microenvironments of advanced human atherosclerotic lesions, intimal fluid becomes acidic. We examined effect an acidic extracellular pH on cholesterol removal (efflux) from primary macrophages. Methods and Results— When efflux acetyl-low-density lipoprotein-loaded macrophages to various acceptors was evaluated at 7.5, 6.5, or 5.5, lower more reduced. The reduction lipid-free apolipoprotein A-I stronger than high-density lipoprotein 2 plasma. Cholesterol every acceptor...

TRAP (thrombospondin-related anonymous protein), localized in the micronemes and on surface of sporozoites notorious malaria parasite Plasmodium, is a key molecule upon infection mammalian host hepatocytes invasion mosquito salivary glands. contains two adhesive domains responsible for cell recognition invasion, known to be essential infectivity. In present paper, we report high-resolution crystal structures A domain Plasmodium falciparum with without bound Mg2+. The structure reveals vWA...

We report the complete sequence of humanCOL9A3 gene that encodes α3 chain heterotrimeric type IX collagen, a member fibril-associated collagens with interrupted triple helices family collagenous proteins. Nucleotide sequencing defined over 23,000 base pairs (bp) and about 3000 bp 5′-flanking sequences. The contains 32 exons. domain exon organization is almost identical to related gene, human COL9A2 gene. However, 2 theCOL9A3 codes for one -Gly-X-Y- triplet less than difference compensated by...

Abstract Calerythrin, a four‐EF‐hand calcium‐binding protein from Saccharopolyspora erythraea , exists in an equilibrium between ordered and less states with slow exchange kinetics when deprived of Ca 2+ at low temperatures, as observed by NMR. As the temperature is raised, signal dispersion NMR spectra reduces, intensity near‐UV CD bands decreases. Yet far‐UV indicate only small decrease amount secondary structure, SAXS data show that no significant change occurs overall size shape protein....

Collagen IX, located on the surface of collagen fibrils, is crucial for cartilage integrity and stability. The N-terminal NC4 domain alpha1(IX) chain probably important in this because it interacts with various macromolecules such as proteoglycans oligomeric matrix protein. At least 17 distinct polypeptides carry an NC4-like unit near their N terminus, but report, describing crystal structure at 1.8-A resolution, represents first atomic level these domains. similar to previously...

Sin3A-associated protein 30-like (SAP30L) is one of the key proteins in a multi-subunit complex involved transcriptional regulation via histone deacetylation. SAP30L, together with highly homologous SAP30 as well other SAP (i.e., SAP25, SAP45, SAP130, and SAP180), an essential component Sin3A corepressor complex, although its actual role has remained elusive. SAP30L thought to function important stabilizing bridging molecule mediate interactions corepressors. been previously shown contain...

Ten UriSed 3 PRO automated microscopes (77 Elektronika, Hungary) were verified for nine HUSLAB laboratories with 160 000 annual urine samples. Particle counting of the primary instrument was against reference visual microscopy 463 specimens, and culture on chromogenic agar plates parallel 396 specimens. Nine secondary instruments compared pairwise instrument. Relative imprecisions to Poisson distribution, R(CV), estimated be 1.0 white blood cell (WBC) 1.5 red (RBC) counts, respectively....

Past attempts to isolate type IX collagen (CIX) from cartilage using limited proteolysis yielded partially degraded material. Recent application of recombinant technology, however, has allowed the preparation intact native CIX. We used murine collagen-induced arthritis model characterize immunologic properties human CIX (rCIX) produced a baculovirus expression system.A panel B10 congenic mice was immunized with rCIX emulsified Freund's complete adjuvant (CFA). The ability induce tolerance...

The complement system is considered the first line of defense against pathogens. Hijacking regulators from blood a common evasion tactic pathogens to inhibit activation on their surfaces. Here, we report hijacking C4b-binding protein (C4bp), regulator classical and lectin pathways activation, by sporozoite (SPZ) stage Plasmodium falciparum parasite. This was shown direct binding radiolabeled purified C4bp live SPZs as well human serum in indirect immunofluorescence assays. Using...