Cloning, Sequencing, and Expression of the Gene from<i>Bacillus circulans</i>That Codes for a Heparinase That Degrades Both Heparin and Heparan Sulfate

Yoshida, Eiichi; Arakawa, S; Matsunaga, Taizo; Toriumi, Shigeki; Tokuyama, Shogo; Morikawa, Kiyoshi; Tahara, Yasutaka

doi:10.1271/bbb.66.1873

Cited by 22 publications

(15 citation statements)

References 31 publications

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“…Glycosaminoglycans remaining in the retentate were collected by reversing the filter and spinning at 12,000 ϫ g, followed by either purification by mini Q H spin column (for PAGE analysis) or incubation with 10-m units of heparin lyase I, II, and III at 35°C for 10 h. The products were recovered by centrifugal filtration using a YM-3 spin column, and the disaccharides were collected in the flow-through and freeze-dried. Cloning, overexpression in Escherichia coli, and purification of the recombinant heparin lyase I (EC 4.2.2.7), heparin lyase II (no EC assigned), and heparin lyase III (EC 4.2.2.8) from Flavobacterium heparinum were all performed as described previously (22,23).…”

Section: Methodsmentioning

confidence: 99%

The Circulating Glycosaminoglycan Signature of Respiratory Failure in Critically Ill Adults

Schmidt

et al. 2014

Journal of Biological Chemistry

131

137

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Background: Endothelial glycocalyx degradation contributes to the pathogenesis of critical illness. Results: Mechanically ventilated subjects exhibited plasma glycocalyx breakdown signatures (glycosaminoglycan fragments) characteristic of direct versus indirect etiologies of respiratory failure. Conclusion: Circulating glycosaminoglycans provide insight into respiratory failure pathophysiology. Significance: This is the first study to characterize circulating glycosaminoglycans during critical illness, offering insight into the mechanisms underlying respiratory failure.

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Section: Methodsmentioning

confidence: 99%

The Circulating Glycosaminoglycan Signature of Respiratory Failure in Critically Ill Adults

Schmidt

et al. 2014

Journal of Biological Chemistry

131

137

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“…AMAC and NaCNBH 3 were purchased from Sigma-Aldrich (St. Louis, USA). The cloning, E. coli expression and purification of the recombinant heparin lyase I (EC 4.2.2.7), heparin lyase II (no EC assigned), and heparin lyase III (EC 4.2.2.8) from F. heparinum were performed in our laboratory as previously described [37–39]. Chinese hamster ovary (CHO)-S cells were grown in suspension culture on CD-CHO medium supplemented with 2% HT (Hypoxanthine/Thymidine mixture, Gibco-Invitrogen) and 8 mM glutamine [40].…”

Section: Methodsmentioning

confidence: 99%

Disaccharide analysis of glycosaminoglycan mixtures by ultra-high-performance liquid chromatography–mass spectrometry

Yang

Chang

Weyers

et al. 2012

Journal of Chromatography A

103

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Glycosaminoglycans are a family of polysaccharides widely distributed in all eukaryotic cells. These polyanionic, linear chain polysaccharides are composed of repeating disaccharide units that are often differentially substituted with sulfo groups. The diversity of glycosaminoglycan structures in cells, tissues and among different organisms reflect their functional an evolutionary importance. Glycosaminoglycan composition and structure also changes in development, aging and in disease progression, making their accurate and reliable analysis a critical, albeit, challenging endeavor. Quantitative disaccharide compositional analysis is one of the primary ways to characterize glycosaminoglycan composition and structure and has a direct relationship with glycosaminoglycan biological functions. In this study, glycosaminoglycan disaccharides, prepared from heparan sulfate/heparin, chondroitin sulfate/dermatan sulfate and neutral hyaluronic acid using multiple polysaccharide lyases, were fluorescently labeled with 2-aminoacridone, fractionated into 17 well-resolved components by reverse-phase ultra-performance liquid chromatography, and analyzed by electrospray ionization mass spectrometry. This analysis was successfully applied to cell, tissue, and biological fluid samples for the picomole level detection of glycosaminoglycan composition and structure.

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“…Unsaturated disaccharides standards of heparin/HS (0S, DUA-GlcNAc; NS, DUA-GlcNS; 6S, DUA-GlcNAc6S; 2S, DUA2S-GlcNAc; 2SNS, DUA2S-GlcNS; NS6S, DUA-GlcNS6S; 2S6S, DUA2S-GlcNAc6S; and TriS, DUA2S-GlcNS6S) were purchased from Seikagaku Corporation ( Japan). Cloning, Escherichia coli expression, and purification of the recombinant heparin lyase I (EC 4.2.2.7), heparin lyase II (no EC assigned), and heparin lyase III (EC 4.2.2.8) from Flavobacterium heparinum were performed in our laboratory as described (Godavarti et al, 1996;Shaya et al, 2006;Yoshida et al, 2002). All other chemicals were of reagent grade.…”

Section: Biological Samples and Materialsmentioning

confidence: 99%

A Structural Analysis of Glycosaminoglycans from Lethal and Nonlethal Breast Cancer Tissues: Toward a Novel Class of Theragnostics for Personalized Medicine in Oncology?

Weyers

Yang

Yoon

et al. 2012

OMICS: A Journal of Integrative Biology

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Cancer is one of the leading noncommunicable diseases that vastly impacts both developed and developing countries. Truly innovative diagnostics that inform disease susceptibility, prognosis, and/or response to treatment (theragnostics) are seriously needed for global public health and personalized medicine for patients with cancer. This study examined the structure and content of glycosaminoglycans (GAGs) in lethal and nonlethal breast cancer tissues from six patients. The glycosaminoglycan content isolated from tissue containing lethal cancer tumors was approximately twice that of other tissues. Molecular weight analysis showed that glycosaminoglycans from cancerous tissue had a longer weight average chain length by an average of five disaccharide units, an increase of approximately 15%. Dissacharide analysis found differences in sulfation patterns between cancerous and normal tissues, as well as sulfation differences in GAG chains isolated from patients with lethal and nonlethal cancer. Specifically, cancerous tissue showed an increase in sulfation at the ''6S'' position of CS chains and an increase in the levels of the HS disaccharide NSCS. Patients with lethal cancer showed a decrease in HS sulfation, with lower levels of ''6S'' and higher levels of the unsulfated ''0S'' disaccharide. Although these findings come from a limited sample size, they indicate that structural changes in GAGs exist between cancerous and noncancerous tissues and between tissues from patients with highly metastatic cancer and cancer that was successfully treated by chemotherapy. Based on these findings, we hypothesize that (1) there are putative changes in the body's construction of GAGs as tissue becomes cancerous; (2) there may be innate structural person-to-person variations in GAG composition that facilitate the metastasis of tumors in some patients when they develop cancer.

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Cloning, Sequencing, and Expression of the Gene fromBacillus circulansThat Codes for a Heparinase That Degrades Both Heparin and Heparan Sulfate

Cited by 22 publications

References 31 publications

The Circulating Glycosaminoglycan Signature of Respiratory Failure in Critically Ill Adults

The Circulating Glycosaminoglycan Signature of Respiratory Failure in Critically Ill Adults

Disaccharide analysis of glycosaminoglycan mixtures by ultra-high-performance liquid chromatography–mass spectrometry

A Structural Analysis of Glycosaminoglycans from Lethal and Nonlethal Breast Cancer Tissues: Toward a Novel Class of Theragnostics for Personalized Medicine in Oncology?

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