Distribution of glycoproteins with β-N-acetylgalactosaminylated N-linked sugar chains among bovine tissues

Sakiyama, Toru; Kabayama, Mineaki; Tomita, Masaru; Nakamura, Jin; Mukai, Hisao; Tomita, Yoshihiro; Furukawa, Kiyoshi

doi:10.1016/s0304-4165(97)00151-7

Cited by 14 publications

(13 citation statements)

References 26 publications

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“…Proteases are used in detergents and cleaning agent for a long time (Sakiyama et al 1998; Showell 1999). The purified metagenomic AS-protease showed maximum activity at pH 10.5 suggesting that it is an alkaline protease (Larsen et al 2006; Moreira 2003).…”

Section: Discussionmentioning

confidence: 99%

Identification and characterization of alkaline serine protease from goat skin surface metagenome

2011

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Metagenomic DNA isolated from goat skin surface was used to construct plasmid DNA library in Escherichia coli DH10B. Recombinant clones were screened for functional protease activity on skim milk agar plates. Upon screening 70,000 clones, a clone carrying recombinant plasmid pSP1 exhibited protease activity. In vitro transposon mutagenesis and sequencing of the insert DNA in this clone revealed an ORF of 1890 bp encoding a protein with 630 amino acids which showed significant sequence homology to the peptidase S8 and S53 subtilisin kexin sedolisin of Shewanella sp. This ORF was cloned in pET30b and expressed in E. coli BL21 (DE3). Although the cloned Alkaline Serine protease (AS-protease) was overexpressed, it was inactive as a result of forming inclusion bodies. After solubilisation, the protease was purified using Ni-NTA chromatography and then refolded properly to retain protease activity. The purified AS-protease with a molecular mass of ~63 kDa required a divalent cation (Co2+ or Mn2+) for its improved activity. The pH and temperature optima for this protease were 10.5 and 42°C respectively.

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Section: Discussionmentioning

confidence: 99%

Identification and characterization of alkaline serine protease from goat skin surface metagenome

2011

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“…3A) and revealed (following electrotransfer of the SDS-PAGE gel to a polyvinylidene difluoride membrane) with horseradish peroxidase-labeled Wisteria floribunda lectin (Fig. 3B), which interacts with oligosaccharides terminating with N-acetylgalactosamine (36). The whole-cell lysates were obtained from the same C. jejuni growth cultures as those used to purify the total RNA for the microarray analysis.…”

Section: Fig 2 Operon Mapping By Rt-pcr Analysis Of Iron-andmentioning

confidence: 99%

Iron Acquisition and Regulation in Campylobacter jejuni

2004

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Iron affects the physiology of bacteria in two different ways: as a micronutrient for bacterial growth and as a catalyst for the formation of hydroxyl radicals. In this study, we used DNA microarrays to identify the C. jejuni genes that have their transcript abundance affected by iron availability. The transcript levels of 647 genes were affected after the addition of iron to iron-limited C. jejuni cells. Several classes of affected genes were revealed within 15 min, including immediate-early response genes as well as those specific to iron acquisition and metabolism. In contrast, only 208 genes were differentially expressed during steady-state experiments comparing iron-rich and iron-limited growth conditions. As expected, genes annotated as being involved in either iron acquisition or oxidative stress defense were downregulated during both time course and steady-state experiments, while genes encoding proteins involved in energy metabolism were upregulated. Because the level of protein glycosylation increased with iron limitation, iron may modulate the level of C. jejuni virulence by affecting the degree of protein glycosylation. Since iron homeostasis has been shown to be Fur regulated in C. jejuni, an isogenic fur mutant was used to define the Fur regulon by transcriptome profiling. A total of 53 genes were Fur regulated, including many genes not previously associated with Fur regulation. A putative Fur binding consensus sequence was identified in the promoter region of most iron-repressed and Fur-regulated genes. Interestingly, a fur mutant was found to be significantly affected in its ability to colonize the gastrointestinal tract of chicks, highlighting the importance of iron homeostasis in vivo. Directed mutagenesis of other genes identified by the microarray analyses allowed the characterization of the ferric enterobactin receptor, previously named CfrA. Chick colonization assays indicated that mutants defective in enterobactin-mediated iron acquisition were unable to colonize the gastrointestinal tract. In addition, a mutation in a receptor (Cj0178) for an uncharacterized iron source also resulted in reduced colonization potential. Overall, this work documents the complex response of C. jejuni to iron availability, describes the genetic network between the Fur and iron regulons, and provides insight regarding the role of iron in C. jejuni colonization in vivo.

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“…From comparison with the constituents of N-linked saccharides, it is clear that Man is a typical monosaccharide for N-glycans. Owing to the discovery of the presence of GalNAc in N-glycans from invertebrates and vertebrates [345][346][347][348][349] already spoken of with respect to the pivotal 4%-sulfated derivative at the termini of glycan antennae of pituitary glycoprotein hormones [37], this sugar should no longer be considered as typical for O-glycans. Biosynthesis of O-glycosidically linked saccharide chains [29,42,339,340] is completely different from that of N-glycans.…”

Section: Mucin-type O-glycansmentioning

confidence: 99%

Eukaryotic glycosylation: whim of nature or multipurpose tool?

Reuter¹,

Gabius²

1999

Cellular and Molecular Life Sciences (CMLS)

214

141

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Protein and lipid glycosylation is a ubiquitous phenomenon. The task of cataloguing the great structural variety of the glycan part has demanded considerable efforts over decades. This patient endeavor was imperative to discern the inherent rules of glycosylation which cannot affirm assumptions on a purely coincidental nature of this type of protein and lipid modification. These results together with theoretical considerations uncover a salient property of oligosaccharides. In comparison with amino acids and nucleotides, monosaccharides excel in their potential to serve as units of hardware for storing biological information. Thus, the view that glycan chains exclusively affect physiochemical properties of the conjugates is indubitably flawed. This original concept has been decisively jolted by the discovery of endogenous receptors (lectins) for distinct glycan epitopes which are as characteristic as a fingerprint or a signature for a certain protein (class) or cell type. Recent evidence documents that these binding proteins are even endowed with the capacity to select distinct low-energy conformers of the often rather flexible oligosaccharides, granting entry to a new level of regulation of ligand affinity by shifting conformer equilibria. The assessment of the details of this recognition by X-ray crystallography, nuclear magnetic resonance spectroscopy, microcalorimetry and custom-made derivatives is supposed to justify a guarded optimism in satisfying the need for innovative drug design in antiadhesion therapy, for example against viral or bacterial infections and unwanted inflammation. This review presents a survey of the structural aspects of glycosylation and of evidence to poignantly endorse the notion that carrier-attached glycan chains can partake in biological information transfer at the level of cell compartments, cells and organs.

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Distribution of glycoproteins with β-N-acetylgalactosaminylated N-linked sugar chains among bovine tissues

Cited by 14 publications

References 26 publications

Identification and characterization of alkaline serine protease from goat skin surface metagenome

Identification and characterization of alkaline serine protease from goat skin surface metagenome

Iron Acquisition and Regulation in Campylobacter jejuni

Eukaryotic glycosylation: whim of nature or multipurpose tool?

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