Bacterial endospores derive much of their longevity and resistance properties from the relative dehydration protoplasts. The spore cortex, a peptidoglycan structure surrounding protoplasm, maintains, is postulated to have role in attaining, protoplast dehydration. A structural modification unique cortex removal all or part peptide side chains majority muramic acid residues conversion 50% lactam. mutation cwlD gene Bacillus subtilis, predicted encode muramoyl-L-alanine amidase, results...

It has been hypothesized that protease-activated receptors may be activated and attenuated by more than one protease. Here, we explore a desensitization mechanism of the PAR1 thrombin receptor anticoagulant proteases provide an explanation to enigma why plasmin/tissue plasminogen activator (t-PA) can both activate deactivate platelets prior treatment. By using soluble N-terminal exodomain (TR78) as model for full-length receptor, were able unambiguously compare cleavage rates specificities...

P-selectin glycoprotein ligand-1 (PSGL-1) is a dimeric membrane mucin on leukocytes that binds selectins. The molecular features of PSGL-1 determine this high affinity binding are unclear. Here we demonstrate the <i>in vitro</i> synthesis novel glycosulfopeptide (GSP-6) modeled after extreme N terminus PSGL-1, which has been predicted to be important for binding. GSP-6 contains three tyrosine sulfate (TyrSO₃) residues and monosialylated, core 2-based <i>O</i>-glycan with sialyl...

P-selectin glycoprotein ligand-1 (PSGL-1) is a mucin on leukocytes that binds to selectins. an N-terminal region of PSGL-1 requires sulfation at least one three clustered tyrosines (TyrSO(3)) and adjacent core-2-based O-glycan expressing sialyl Lewis x (C2-O-sLe(x)). We synthesized glycosulfopeptides (GSPs) modeled after this explore the roles individual TyrSO(3) residues, placement C2-O-sLe(x) relative TyrSO(3), contributions fucose sialic acid C2-O-sLe(x), function peptide sequence for...

ABSTRACT Of the toxins produced by Bacillus cereus , emetic toxin is likely most dangerous but, due to lack of a suitable assay, least well known. In this paper, new, sensitive, inexpensive, and rapid bioassay for detection B. described. The assay based on loss motility boar spermatozoa upon 24 h exposure extracts strains or contaminated food. paralyzed exhibited swollen mitochondria, but no depletion cellular ATP damage plasma membrane integrity was observed. Analysis purified electrospray...

Peptidoglycan was prepared from purified Bacillus subtilis spores of wild-type and several mutant strains. Digestion with muramidase resulted in cleavage the glycosidic bonds adjacent to muramic acid replaced by peptide or alanine side chains but not lactam. Reduction resulting muropeptides allowed their separation reversed-phase high-pressure liquid chromatography. The structures 20 were determined amino sugar analysis matrix-assisted laser desorption/ionization time-of-flight mass...

Complex carbohydrate structures, glycans, are essential components of glycoproteins, glycolipids, and proteoglycans. While individual glycan structures including the SSEA Tra antigens already used to define undifferentiated human embryonic stem cells (hESC), whole spectrum cell glycans has remained unknown. We undertook a global study asparagine-linked glycoprotein (N-glycans) hESC their differentiated progeny using MALDI-TOF mass spectrometric NMR spectroscopic profiling. Structural...

The P-selectin counterreceptor PSGL-1 is covalently modified by mono alpha2,3-sialylated, multiply alpha1,3-fucosylated polylactosamines. These glycans are required for the adhesive interactions that allow this adhesion receptor-counterreceptor pair to facilitate leukocyte extravasation. To begin understand biosynthesis of these glycans, we have characterized acceptor and site specificities two granulocyte alpha1,3-fucosyltransferases, Fuc-TIV Fuc-TVII, using recombinant forms enzymes a...

ABSTRACT Actinomycete isolates from indoor air and dust in water-damaged schools children’s day care centers were tested for toxicity by using boar spermatozoa as an indicator. Toxicity was detected extracts of four strains which caused a loss sperm motility, the 50% effective concentrations (EC 50 ) 10 to 63 ng (dry weight) ml extended semen −1 . The identified Streptomyces griseus 16S ribosomal DNA chemotaxonomic methods. S. had similar effects on cells, including motility swelling...

Nisin produced by Lactococcus lactis subsp. is a 34-residue antibacterial polypeptide and belongs to group of post-translationally modified peptides, lantibiotics, with dehydrated residues cyclic amino acids, lanthionines. These modifications are supposed be made enzymes encoded lanB lanC genes, found only in biosynthetic operons encoding lantibiotics. To analyse the extent modification, His-tagged nisin precursors were expressed nisB nisC mutant strains. The purified from cytoplasm cells,...

Activation of plasminogen (plg) to plasmin by the staphylococcal activator, staphylokinase (SAK), is effectively regulated circulating inhibitor, α 2 ‐antiplasmin (α AP). Here it demonstrated that intact Staphylococcus aureus cells and solubilized cell wall proteins not only protected SAK‐promoted plg activation against inhibitory effect AP but also enhanced activation. The findings suggest surface‐associated SAK may have an important physiological function in helping staphylococci tissue...

Toxins from three Bacillus licheniformis strains connected to a fatal food poisoning were isolated and their structures elucidated. purified methanol extracts of the B. biomass using boar sperm cells as toxicity indicator. The HPLC toxins showed protonated masses m/z 1007, 1021 1035 in MALDI-TOF-MS. different origins contained same components which each had amino-acid residues L-Gln, L-Leu, D-Leu, L-Val, L-Asp, D-Leu L-Ile that order. identified lichenysin A, cyclic lactonic heptalipopeptide...

VP4, the major structural protein of haloarchaeal pleomorphic virus, HRPV-1, is glycosylated. To define glycan structure attached to this protein, oligosaccharides released by β-elimination were analysed mass spectrometry and nuclear magnetic resonance spectroscopy. Such analyses showed that VP4-derived a pentasaccharide comprising glucose, glucuronic acid, mannose, sulphated acid terminal 5-N-formyl-legionaminic residue. This first observation legionaminic sialic acid-like sugar, in an...

Antibody-drug conjugates (ADCs) offer a combination of antibody therapy and specific delivery potent small-molecule payloads to target cells. The properties the ADC molecule are determined by balance its components. efficacy payload component increases with higher drug-to-antibody ratio (DAR), while homogeneous DAR = 8 ADCs easily prepared conjugation four accessible hinge cystines. However, use hydrophobic has permitted only 2–4, due poor pharmacokinetics aggregation problems. Here, we...

The prevalence of acute respiratory infections and their impact on quality life underlies the need for efficacious solutions that are safe, sustainable economically viable. Polysaccharides in several (traditional) plant extracts have been shown to be immunostimulatory, some studies suggest beneficial effects against infections. aim this study was (i) identify active polysaccharide constituents from affordable renewable crops (bell pepper carrot) using activity-guided fractionation, (ii)...

Background: We have previously identified patatin (Sol t 1) of potato tubers as a major food allergen among atopic children. In addition to Sol 1, concomitant IgE binding other, then unidentified, proteins was observed. Methods: Purification and identification the putative allergens were done by both standard advanced methods protein chemistry. The patient series comprised 39 children with positive skin prick test (SPT) raw potato. Immunoblotting ELISA used examine IgE‐binding ability...

Abstract Adenosylmethionine decarboxylase (AdoMetDC), a key enzyme in the biosynthesis of polyamines, is often up-regulated cancers. We have demonstrated previously that overexpression AdoMetDC alone sufficient to transform NIH 3T3 cells and induce highly invasive tumors nude mice. Here, we studied transformation-specific alterations gene expression induced by using cDNA microarray two-dimensional electrophoresis technologies. specifically tried identify secreted proteins contributing high...

Matrix metalloproteinase-1 (MMP-1) is a collagenolytic metalloproteinase capable of cleaving native triple-helical forms several collagen subtypes, as well number non-collagenous substrates. The role MMP-1 in various diseases affecting the connective tissue characterized. secreted both glycosylated and unglycosylated species, two have been shown to be identical with respect substrate specificity, specific activity inhibitory profile. No function for glycan moiety enzyme has ascribed date. In...

Fucosylation of glycans on glycoproteins and -lipids requires the enzymatic activity relevant fucosyltransferases GDP-L-fucose as donor. Due to biological importance fucosylated glycans, a readily accessible source would be required. Here we describe construction stable recombinant S.cerevisiae strain expressing E.coli genes gmd wcaG encoding two enzymes, GDP-mannose-4,6-dehydratase (GMD) GDP-4-keto-6-deoxy-D-mannose-3,5-epimerase/4-reductase (GMER(FX)) respectively, needed convert...

We have previously isolated and partially characterised the components of a highly purified interferon‐α (IFN‐α) preparation produced by Sendai‐virus‐induced human peripheral blood leukocytes. Nine IFN‐α species were identified, two these found to be glycosylated [Nyman, T. A., Tölö, H., Parkkinen, J. &amp; Kalkkinen, N. (1998) Identification nine subtypes leukocytes, Biochem. 329 , 295−302]. Here, we N‐linked oligosaccharides IFN‐α14c O‐linked chains IFN‐α2b, glycans electrospray tandem...

A novel approach to localize and reconstruct conformational IgE-binding epitope regions of hevein (Hev b6.02), a major natural rubber latex allergen, is described. An antimicrobial protein (AMP) from the amaranth <i>Amaranthus caudatus</i> was used as an immunologically non-IgE-binding adaptor molecule which terminal or central parts were fused. Hevein AMP share structurally identical core region but have different N-terminal C-terminal regions. Only 1 16 hevein-allergic patients showed weak...

Antibody-drug conjugates (ADCs) are promising state-of-the-art biopharmaceutical drugs for selective drug-delivery applications and the treatment of diseases such as cancer. The idea behind ADC technology is remarkable it combines highly targeting capacity monoclonal antibodies with cancer-killing ability potent cytotoxic agents. continuous development improved ADCs requires systematic studies on nature effects warhead modification. Recently, we focused hydrophilic modification monomethyl...