Bacillus circulans chitinase A1 (ChiA1) has a deep substrate-binding cleft on top of its (beta/alpha)8-barrel catalytic domain and an interaction between the aromatic residues in this bound oligosaccharide been suggested. To study roles these residues, especially crystalline-chitin hydrolysis, site-directed mutagenesis was carried out. Y56A W53A mutations at subsites -5 -3, respectively, selectively decreased hydrolysing activity against highly crystalline beta-chitin. W164A W285A +1 +2,...

The rice class I chitinase OsChia1b, also referred to as RCC2 or Cht-2, is composed of an N-terminal chitin-binding domain (ChBD) and a C-terminal catalytic (CatD), which are connected by proline- threonine-rich linker peptide. Because the ability inhibit fungal growth, OsChia1b gene has been used produce transgenic plants with enhanced disease resistance. As initial step toward elucidating mechanism hydrolytic action antifungal activity, full-length structure was analyzed X-ray...

Significance The Red Queen hypothesis proposes that host defense genes evolve to counter the adverse effects of rapidly evolving invasive viruses. Although 3D structures host–viral protein complexes have provided great insights into molecular conflicts between them, a single structure represents only an evolutionary snapshot. Here we present atomic details step-by-step arms race tomato mosaic virus replication and inhibitor Tm-1, in which recognition viral molecule, adaptive evasion...

Chitinase C from Streptomyces griseus HUT6037 was discovered as the first bacterial chitinase in family 19 other than chitinases found higher plants. comprises two domains: a chitin-binding domain (ChBDChiC) for attachment to chitin and chitin-catalytic digesting chitin. The structure of ChBDChiC determined by means 13C-, 15N-, 1H-resonance nuclear magnetic resonance (NMR) spectroscopy. conformation its backbone comprised β-sheets composed three antiparallel β-strands, respectively, this...

Fusobacterium nucleatum produces a large amount of the toxic metabolite hydrogen sulfide in oral cavity. Here, we report molecular basis F. H 2 S production, which is associated with two different enzymes: previously reported Cdl (Fn1220) and newly identified Lcd (Fn0625). SDS-PAGE analysis activity staining revealed that crude enzyme extracts from ATCC 25586 contained three major S-producing proteins. Two proteins low masses migrated similarly to purified Fn0625 Fn1220. Their kinetic values...

Functional investigation of the proposed dehydratase domain ATX, a 6-methylsalicylic acid synthase from Aspergillus terreus, revealed that is not involved in dehydration β-hydroxytriketide intermediate tethered on acyl carrier protein but catalyzes thioester hydrolysis to release product protein. Thus, we renamed this hydrolase (TH) domain. The bound TH mutant H972A formed presence NADPH was released as by both intact ATX and THID (a 541-amino region containing its downstream) protein,...

In the three-dimensional structure of a rice class I chitinase (OsChia1b) determined recently, loop (loop II) is located at end substrate-binding cleft, and thus suggested to be involved in substrate binding. order test this assumption, deletion II region from catalytic domain OsChia1b replacement Trp159 with Ala were carried out. The W159A mutation increased hydrolytic activity not only towards (GlcNAc)6 but also polysaccharide substrates. Similar results obtained for kcat/Km values...

A third enzyme that produces hydrogen sulfide from L-cysteine was identified in Fusobacterium nucleatum subsp. nucleatum. The fn1055 gene cloned a cosmid library constructed with genomic DNA of F. ATCC 25586. Despite the database annotation product is cysteine synthase, reverse-phase HPLC revealed no produced vitro by purified Fn1055 protein; however, did produce L-serine. In addition, auxotroph, Escherichia coli NK3, transformed plasmid containing not grow without cysteine, which further...

Abstract Hydrogen sulfide (H 2 S) is a causative agent of oral malodor and may play an important role in the pathogenicity bacteria such as Streptococcus anginosus . In this microorganism, H S production associated with βC‐S lyase (Lcd) encoded by lcd gene, which pyridoxal 5′‐phosphate (PLP)‐dependent enzyme that catalyzes α,β‐elimination sulfur‐containing amino acids. When Lcd acts on L ‐cysteine, produced along pyruvate ammonia. To understand S‐producing mechanism detail, we determined...

Hydrogen sulfide (H2S) plays important roles in the pathogenesis of periodontitis. Oral pathogens typically produce H2S from l-cysteine addition to pyruvate and [Formula: see text] However, fn1055 Fusobacterium nucleatum subsp. ATCC 25586 encodes a pyridoxal 5'-phosphate (PLP)-dependent enzyme that catalyzes production l-serine H2O, an unusual cysteine (hydroxyl) lyase reaction (β-replacement reaction). To reveal mechanism, crystal structure substrate-free Fn1055 was determined. Based on...

Gibberellins (GAs) are phytohormones regulating various developmental processes in plants. In rice, the initial GA-signaling events involve binding of a GA to soluble receptor protein, GID1. Although X-ray structures for certain GID1/GA complexes have recently been determined, an examination does not fully clarify how GID1s discriminate among different GAs. Herein, we present study aimed at defining types forces important via combination isothermal titration calorimetry (ITC) and...

Acetyl phosphate (AcP) is generally produced from acetyl coenzyme A by phosphotransacetylase (Pta), and subsequent reaction with ADP, catalyzed acetate kinase (Ack), produces ATP. The mechanism of ATP production in Porphyromonas gingivalis poorly understood. aim this study was to explore the molecular basis Pta-Ack pathway microorganism. Pta Ack P. ATCC 33277 were enzymatically structurally characterized. Structural mutational analyses suggest that a dimer two substrate-binding sites each...

Gibberellins (GAs) are phytohormones that regulate various developmental processes in plants. The initial GA signalling events involve the binding of a to soluble receptor protein GID1, followed by complex negative transcriptional regulator signaling, DELLA protein. Although X-ray structures for certain Arabidopsis GID1/GA/DELLA complexes have previously been determined, examination these did not fully clarify how recognizes and binds GID1/GA complex. Herein, we present study aimed at...

To determine the properties and structure of OsChia1b, a family 19 chitinase from Oryza sativa L. cv. Nipponbare (japonica ssp.), recombinant OsChia1b was produced in Esherichia coli cells purified to homogeneity by chitin affinity column chromatography. highly active against soluble chitinous substrate, but not crystalline chitin, clearly inhibited hyphal extension Trichoderma reesei.

X-linked adrenal hypoplasia congenita (AHC) is a rare disorder characterized by primary insufficiency and hypogonadotropic hypogonadism (HHG) caused mutations of the NR0B1/DAX1 gene. We aimed to search for presence any gene in referred patient further characterize phenotypes identified mutation.Herein, we report Japanese with novel missense mutation resulting adult-onset AHC HHG. The was diffuse skin pigmentation at 28 years age, presented partial had undiagnosed incomplete Urological...

We report here on crystallization and preliminary X-ray analysis of plant class I chitinase from rice (OsChia1b). Similar single crystals full-length OsChia1b were obtained under two independent conditions. The grown these conditions diffracted up to 2.1 2.5 Å resolution, respectively, at a synchrotron beamline, found belong the tetragonal space group P43212. Keywords: chitinase, (oryza sativa l. japonica), crystallization, x-ray analysis, protease inhibitor complex, specificity, elastase

Tm-1, an inhibitor protein of Tomato mosaic virus RNA replication, contains two conserved domains: uncharacterized domain at its N-terminus and a TIM-barrel-like C-terminus. The N-terminal Tm-1 has inhibitory activity three-dimensional structure not been determined. Here, the crystallization preliminary X-ray diffraction are reported. A three-wavelength MAD data set was collected from selenomethionine-labelled crystal processed to 2.7 Å resolution. belonged triclinic space group P1, with...

Hydrogen sulfide produced by oral bacteria is responsible for malodour. Two homologous hydrogen sulfide-producing enzymes, Fn1220 and Cdl, from Fusobacterium nucleatum (which actively produces sulfide) were overproduced, purified crystallized. X-ray diffraction data collected the crystals using a synchrotron-radiation source. The crystal belonged to tetragonal space group P4(1)2(1)2 or P4(3)2(1)2 (unit-cell parameters a=b=116.8, c=99.2 Å) Cdl monoclinic P2(1) a=84.9, b=70.9, c=87.6 Å, β=90.3°).

A 16 kDa buckwheat protein (BWp16) is a major allergen responsible for immediate hypersensitivity reactions including anaphylaxis. deletion mutant of BWp16 (rBWp16ΔN) was overproduced and purified shown to be immunologically active. three-wavelength MAD data set collected from crystal selenomethionine-labelled rBWp16ΔN. The belonged the triclinic space group P1, with unit-cell parameters = 28.39, b 31.54, c 32.20 Å, α 111.92, β 108.91, γ 98.74°. One monomer expected present in asymmetric...

A new three-dimensional graphics program, SaxsMDView, is described. The program performs a graphical representation for protein molecules along with the force vector (or potential) applying to each atom. displayed object can be rotated and translated in arbitrary directions by interactive mouse manipulation. While SaxsMDView was originally intended visualize result of SAXS_MD, previously developed based on restrained molecular dynamics small-angle X-ray scattering constraints, it also useful...