• 대한유전성대사질환학회지
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• 제14권2호
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• pp.178-181
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• 2014

헌터증후군(뮤코다당증 II형)은 글리코사미노글리칸의 분해를 촉매하는 효소인 iduronate-2-sulfatase 결핍에 의해 조직이나 기관의 세포 내 리소좀에 heparin sulfate와 dermatan sulfate 등의 전구물질이 축적되어 퇴행성 병변을 일으키는 유전 질환이다. 현재 효소보충요법을 통해 증상의 호전 및 질병의 진행을 지연시키는 치료가 가능하나, 중추신경계 증상이 발현된 경우 치료가 어려운 한계가 있어, 무엇보다 조기에 의심하고 진단하여 치료를 시작하는 것이 중요하다. 따라서 어린연령에 진단된 환아들의 임상적 특징에 대해 이해하는 것이 필요하며, 이에 저자들은 2.5세의 어린 연령에 진단된 환아를 경험하여 이를 보고하는 바이다.

• Journal of Microbiology and Biotechnology
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• 제23권5호
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• pp.689-698
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• 2013

The production and characterization of an active recombinant N-acetylgalactosamine-6-sulfate sulfatase (GALNS) in Escherichia coli BL21(DE3) has been previously reported. In this study, the effect of the signal peptide (SP), inducer concentration, process scale, and operational mode (batch and semi-continuous) on GALNS production were evaluated. When native SP was presented, higher enzyme activity levels were observed in both soluble and inclusion bodies fractions, and its removal had a significant impact on enzyme activation. At shake scale, the optimal IPTG concentrations were 0.5 and 1.5 mM for the strains with and without SP, respectively, whereas at bench scale, the highest enzyme activities were observed with 1.5 mM IPTG for both strains. Noteworthy, enzyme activity in the culture media was only detected when SP was presented and the culture was carried out under semi-continuous mode. We showed for the first time that the mechanism that in prokaryotes recognizes the SP to mediate sulfatase activation can also recognize a eukaryotic SP, favoring the activation of the enzyme, and could also favor the secretion of the recombinant protein. These results offer significant information for scaling-up the production of human sulfatases in E. coli.

• Clinical and Experimental Reproductive Medicine
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• 제46권4호
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• pp.206-210
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• 2019

Mucopolysaccharidosis type II (MPS II) is a rare X-linked recessive lysosomal storage disease caused by mutation of the iduronate-2-sulfatase gene. The mutation results in iduronate-2-sulfatase deficiency, which causes the progressive accumulation of heparan sulfate and dermatan sulfate in cellular lysosomes. The phenotype, age of onset, and symptoms of MPS II vary; accordingly, the disease can be classified into either the early-onset type or the late-onset type, depending on the age of onset and the severity of the symptoms. In patients with severe MPS II, symptoms typically first appear between 2 and 5 years of age. Patients with severe MPS II usually die in the second decade of life although some patients with less severe disease have survived into their fifth or sixth decade. Here, we report the establishment of a preimplantation genetic diagnosis (PGD) strategy using multiplex nested polymerase chain reaction, direct sequencing, and linkage analysis. Unaffected embryos were selected via the diagnosis of a single blastomere, and a healthy boy was delivered by a female carrier of MPS II. This is the first successful application of PGD in a patient with MPS II in Korea.

• Journal of Interdisciplinary Genomics
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• 제1권1호
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• pp.6-9
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• 2019

In this report, the phenotypes of three patients from two families with mucopolysaccharidosis type II (MPS II) are compared: a novel variant and recombinant variant of IDS gene. The results of urine in patients showed a pronounced increase in glycosaminoglycan excretion with decreased iduronate-2-sulfatase enzyme activity in leukocyte, leading to a diagnosis of MPS II. A patient has a novel variant with 1 bp small insertion, c.1224_1225insC in exon 9, which caused frameshifts with a premature stop codon, and two patients have a recombination variant, c.418+495_1006+1304del, leading to the loss of exons 4, 5, 6, and 7 in genomic DNA, which is relatively common in Korean patients. They had different phenotypes even in the same mutation. The patients have now been enzyme replacement therapy with a significant decrease in glycosaminoglycan excretion. Further study on residual enzyme activity, as well as experience with more cases, may shed light on the relationship between phenotypes in MPS II and gene mutations.

• Clinical and Experimental Pediatrics
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• 제55권3호
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• pp.88-92
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• 2012

Purpose: Mucopolysaccharidosis type II (MPS II or Hunter syndrome) is a rare lysosomal storage disorder caused by iduronate-2-sulfatase (IDS) deficiency. MPS II causes a wide phenotypic spectrum of symptoms ranging from mild to severe. IDS activity, which is measured in leukocyte pellets or fibroblasts, was reported to be related to clinical phenotype by Sukegawa-Hayasaka et al. Measurement of residual plasma IDS activity using a fluorometric assay is simpler than conventional measurements using skin fibroblasts or peripheral blood mononuclear cells. This is the first study to describe the relationship between plasma IDS activity and clinical phenotype of MPS II. Methods: We hypothesized that residual plasma IDS activity is related to clinical phenotype. We classified 43 Hunter syndrome patients as having attenuated or severe disease types based on clinical characteristics, especially intellectual and cognitive status. There were 27 patients with the severe type and 16 with the attenuated type. Plasma IDS activity was measured by a fluorometric enzyme assay using 4-methylumbelliferyl- ${\alpha}$-iduronate 2-sulphate. Results: Plasma IDS activity in patients with the severe type was significantly lower than that in patients with the attenuated type ($p$=0.006). The optimal cut-off value of plasma IDS activity for distinguishing the severe type from the attenuated type was 0.63 $nmol{\cdot}4hr^{-1}{\cdot}mL^{-1}$. This value had 88.2% sensitivity, 65.4% specificity, and an area under receiver-operator characteristics (ROC) curve of 0.768 (ROC curve analysis; $p$=0.003). Conclusion: These results show that the mild phenotype may be related to residual lysosomal enzyme activity.

• 대한유전성대사질환학회지
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• 제15권2호
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• pp.87-92
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• 2015

뮤코다당증(mucopolysaccharidosis)은 글리코사미노글리칸(glycosaminoglycans)의 분해에 필요한 리소좀 효소의 결함으로 인해 야기되는 질병으로 글리코사미노글리칸의 대사 산물이 세포의 리소좀 내에 축적되어 세포, 조직 그리고 기관의 기능 이상을 초래해 신체적, 신경학적인 퇴행을 보이며, 심한 경우 조기에 사망하게 되는 다양한 임상양상을 보이는 질환이다. 뮤코다당증 가운데 가장 높은 비율을 차지하는 헌터증후군(뮤코다당증 제2형)은 조기에 진단하여 효소보충요법을 시행하는 것이 중요하다. 본 증례는 언어발달지연과 등과 엉덩이에 몽고반점, 간비대, 두껍고 거친 피부가 있었으며 과성장된 신체 검진소견을 보였던 환아에서, 뇌자기공명영상 검사 결과에서 뇌교량체에 다수의 낭종, 백색질에 비정상 신호 증가 병변들, 미만성 뇌수축 소견을 보여 헌터증후군을 의심하였으며 효소검사 결과를 통해 확진하였다. 저자들은 언어발달지연을 주소로 내원한 환아에게 시행한 뇌자기공명영상에서 조기에 헌터증후군을 의심하여 효소검사를 통해 확진을 할 수 있었던 증례를 경험하였기에 이를 보고하는 바이다.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1992년도 제1회 신약개발 연구발표회 초록집
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• pp.23-23
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• 1992

횐 쥐에서 sulfotransferase 양성균주를 검색한 결과 호기성 조건보다는 혐기성 조건의 균이 많은 것으로 나타났다. 이러한 곁과는 장내미생물의 대부분이 혐기성균이라는 것과도 일치하는 것이다. 검색용 배지에서 형광을 나타내는 Sulfotransferase를 생산하는 4개 균주에서 가장 활성이 높은 k-36균주에 대하여 동정하였다. K-36 균주는 혐기적 및 호기적 양조건에서 잘 자라는 통성혐기성균이고, Gas를 생산하며 포자를 생성하지 않는 Gram음성의 간균이었다. 이것으로 보아 이균은 Enterobacteriacease의 Klebsiella oxytoca 로 동정했다. K-36에서 분리한 Sulfotransferase의 반응양식은 p-nitrophenylsulfate와 p-nitrophenol이 반응시간과 함께 동량씩 감소하여 반응 생성물인 phenylsulfate와 p-nitrophenol을 생성시켰으며 sulfatase 반응은 진행되지 않았다.

• Applied Biological Chemistry
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• 제20권1호
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• pp.136-141
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• 1977

돼지 백혈구(白血球) Iysosome의 n-butanol 추출물(抽出物)로 부터 DEAE-cellulose, Sephadex c-200 column chromatography 및 Polyacrylamide gel electrophoresis 로서 acid phosphatase, aryl sulfatase, ${\beta}-glucuronidase$로서 및 cathepsin D를 분리(分離)정제하고 그 성질을 조사하였다.

• Journal of Microbiology and Biotechnology
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• 제25권11호
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• pp.1894-1901
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• 2015

A novel sulfatase gene, ary423 (1,536 bp ORF), encoding a protein of 511 amino acids with a calculated molecular mass of 56 kDa, was identified from Flammeovirga pacifica, which was isolated from deep-sea sediments of west Pacific Ocean. Amino acid sequence analysis revealed that Ary423 possessed a conserved C-X-A-X-R motif, which was recognized as the sulfatase signature. Phylogenetic analysis suggested that Ary423 belonged to arylsulfatases. After heterologous expression in Escherichia coli cells, the recombinant Ary423 was purified with a Ni⁺ affinity column, and was shown to be highly active at a broad range of temperatures from 30° to 70℃, with maximum activity at 40℃. Furthermore, recombinant Ary423 retained more than 70% and 40% of its maximum activity after 12 h of incubation at 50℃ and 60℃, respectively, exhibiting good thermostability at high temperatures. The optimal pH for Ary423 was determined to be 8.0 and the activity of Ary423 could be slightly enhanced by Mg²⁺. The recombinant enzyme could hydrolyze sulfate ester bonds in p-nitrophenyl sulfate (NPS) and Asparagus crude polysaccharides with a specific activity of 64.8 U/mg and 25.4 U/mg, respectively. These favorable properties could make Ary423 attractive for application in the desulfating process of agar production.

• Archives of Pharmacal Research
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• 제26권4호
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• pp.258-263
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• 2003

Colon-specific delivery of glucocorticoids is highly desirable for the efficient treatment of inflammatory bowel disease. We synthesized prednisolone 21-sulfate sodium (PDS) as a colon-specific prodrug of prednisolone (PD) and investigated its properties using rats as test animals. We expected that introduction of sulfate ester as a sodium salt might increase the hydrophilicity and restrict the absorption in the GI tract. If PDS is stable and nonabsorbable in the upper intestine, it will be delivered to the colon as an intact form, where it hydrolyze by the sulfatase to release PD. Compared with PD, the solubility of PDS increased and the apparent partition coefficient decreased greatly. PDS was stable on incubation with pH 1.2 and 6.8 buffer solutions and with the contents of the stomach and small intestine. On incubation with the cecal contents, PDS decreased to 9.6％ of the dose in 10 h producing PD. The amount of PD increased to give a maximum 54％ of the dose and decreased. As a control, when PD was incubated with the cecal contents, it decreased to 29％ of the dose in 8 h, which implied that reduction of PD proceeded under such conditions. These results suggested that hydrolysis of PDS took place to produce and accumulate PD, which decreased by reduction as the incubation period extended. Our results suggested that PDS can be a promising colon-specific prodrug of PD, and sulfate ester group might serve as a potential colon-specific promoiety, especially for the drugs which are resistant to reduction in the colon.