• Title/Summary/Keyword: Side chain cleavage

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The Conversion of Lithocholic Acid into 5$\beta$-Androstan-3, 17-dione in the Cell-free System of Mycobacterium sp. NRRL B-3805

  • Lee, Kang-Man;Park, Hye-Kyung
    • Archives of Pharmacal Research
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    • v.14 no.3
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    • pp.261-265
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    • 1991
  • In a microbial cell-free extract system, side chain cleavage on various sterols and steroids was tested. The cell-free extracts of Mycobacterium sp. NRRL B-3805 showed the side chain cleavage activity on lithocholic acid to form 5$\beta$-androstan-3.17-dione. The properties of the activity were examined.

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Cholesterol side-chain cleavage enzyme deficiency caused by a novel homozygous variant in P450 sidechain cleavage enzyme gene (CYP11A1) in a 46,XX Korean girl

  • Ye Ji Kim;Sun Cho;Hwa Young Kim;Young Hwa Jung;Jung Min Ko;Chang Won Choi;Jaehyun Kim
    • Journal of Genetic Medicine
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    • v.20 no.1
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    • pp.25-29
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    • 2023
  • The CYP11A1 gene encodes for the cholesterol side-chain cleavage enzyme (P450scc), which initiates steroid hormone biosynthesis. Defective P450scc activity results in severe glucocorticoid and mineralocorticoid deficiencies. We describe a case of P450scc deficiency due to a novel homozygous CYP11A1 variant inherited from the mother with a possibility of uniparental disomy (UPD). The patient was a female, had no family history of endocrine disease, and showed adrenal insufficiency at 13 days of age. Hormonal analysis with an adrenocorticotropic hormone stimulation test showed both glucocorticoid and mineralocorticoid deficiencies, presumed to be a defect of the early stage of steroidogenesis. Exome sequencing reported a novel homozygous frameshift variant of CYP11A1 (c.284_285del, p.Asn95Serfs*10), which was inherited from the mother. Additionally, homozygosity in 15q22.31q26.2, which included CYP11A1, was identified using a chromosomal microarray. It was suggested that the possibility of maternal UPD was involved as the cause of a P450scc deficiency by unmasking the maternally derived affected allele. To our understanding, P450scc deficiency associated with UPD encompassing CYP11A1 had not been reported in Korea before. Genetic analysis can help diagnose rare causes of primary adrenal insufficiency, including P450scc deficiency.

Interaction Models of Substrate Peptides and β-Secretase Studied by NMR Spectroscopy and Molecular Dynamics Simulation

  • Lee, Jee-Young;Lee, Sung-Ah;Kim, Jin-Kyoung;Chae, Chi-Bom;Kim, Yangmee
    • Molecules and Cells
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    • v.27 no.6
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    • pp.651-656
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    • 2009
  • The formation of ${\beta}$-amyloid peptide ($A{\beta}$) is initiated from cleavage of amyloid precursor protein (APP) by a family of protease, ${\alpha}$-, ${\beta}$-, and ${\gamma}$-secretase. Sub W, a substrate peptide, consists of 10 amino acids, which are adjacent to the ${\beta}$-cleavage site of wild-type APP, and Sub M is Swedish mutant with double mutations on the left side of the ${\beta}$-cleavage site of APP. Sub W is a normal product of the metabolism of APP in the secretary pathway. Sub M is known to increase the efficiency of ${\beta}$-secretase activity, resulting in a more specific binding model compared to Sub W. Three-dimensional structures of Sub W and Sub M were studied by CD and NMR spectroscopy in water solution. On the basis of these structures, interaction models of ${\beta}$-secretase and substrate peptides were determined by molecular dynamics simulation. Four hydrogen bonds and one water-mediated interaction were formed in the docking models. In particular, the hydrogen bonding network of Sub M-BACE formed spread over the broad region of the active site of ${\beta}$-secretase (P5-P3'), and the side chain of P2- Asn formed a hydrogen bond specifically with the side chain of Arg235. These are more favorable to the cleavage of Sub M by ${\beta}$-secretase than Sub W. The two substrate peptides showed different tendency to bind to ${\beta}$-secretase and this information may useful for drug development to treat and prevent Alzheimer's disease.

Mechanisms of Lignin Biodegradation by Ligninase, Phanerochaete Chrysosporium Burds

  • Hwang, Byung-Ho
    • Journal of Forest and Environmental Science
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    • v.6 no.1
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    • pp.45-60
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    • 1989
  • 리그닌 생분해의 분해경로 및 매카니즘에 관한 연구가 최근 Kirk와 Higuchi 등에 의하여 활발히 연구되고 있다. 특히, Phanerochaete chrysosporium이 생산해내는 Lignlnase를 이용하여 매우 가치있는 연구 결과를 얻고 있다. 본 총설에서는 Kirk와 Higuchi의 허가를 얻어서 그들의 논문을 중심으로 리그닌의 중요한 결합 양식 별로 즉, ${\beta}$-O-4, ${\beta}$-5, ${\beta}$-1, ${\beta}$-6, 5-5 등의 결합 모델 화합물들의 분해경로 및 매카니즘에 관하여 조사 정리하였다.

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Characterization of the Fragmentation Pattern of Peptide from Tandem Mass Spectra

  • Ramachandran, Sangeetha;Thomas, Tessamma
    • Mass Spectrometry Letters
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    • v.10 no.2
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    • pp.50-55
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    • 2019
  • The fragmentation statistics of ion trap CID (Collision-Induced Dissociation) spectra using 87,661 tandem mass spectra of doubly charged tryptic peptides are analyzed here. In contrast to the usual method of using intensity information, the frequency of occurrence of fragment ions, with respect to the position of the cleavage site and the residues at these sites is studied in this paper. The analysis shows that the frequency of occurrence of fragment ion peaks is more towards the middle of the peptide than its ends. It was noted that amino acid with an aromatic and basic side chain at N- & C- terminal end of the peptide stimulates more peaks at the lower end of the spectrum. The residue pair effect was shown when the amide bond occurs between acidic and basic residues. The fragmentation at these sites (D/E-H/R/K) stimulates the generation of the y-ion peak. Also, the cleavage site H-H/R/K stimulates the generation of b-ions. K-P environment in the peptide sequence has more tendency to generate y-ions than b-ions. Statistical analysis helps in the visualization of the CID fragmentation pattern. Cleavage pattern along the length of the peptide and the residue pair effects, enhance the knowledge of fragmentation behavior, which is useful for the better interpretation of tandem mass spectra.

Moieties of Complement iC3b Recognized by the I-domain of Integrin αXβ2

  • Choi, Jeongsuk;Buyannemekh, Dolgorsuren;Nham, Sang-Uk
    • Molecules and Cells
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    • v.43 no.12
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    • pp.1023-1034
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    • 2020
  • Complement fragment iC3b serves as a major opsonin for facilitating phagocytosis via its interaction with complement receptors CR3 and CR4, also known by their leukocyte integrin family names, αMβ2 and αXβ2, respectively. Although there is general agreement that iC3b binds to the αM and αX I-domains of the respective β2-integrins, much less is known regarding the regions of iC3b contributing to the αX I-domain binding. In this study, using recombinant αX I-domain, as well as recombinant fragments of iC3b as candidate binding partners, we have identified two distinct binding moieties of iC3b for the αX I-domain. They are the C3 convertase-generated N-terminal segment of the C3b α'-chain (α'NT) and the factor I cleavage-generated N-terminal segment in the CUBf region of α-chain. Additionally, we have found that the CUBf segment is a novel binding moiety of iC3b for the αM I-domain. The CUBf segment shows about a 2-fold higher binding activity than the α'NT for αX I-domain. We also have shown the involvement of crucial acidic residues on the iC3b side of the interface and basic residues on the I-domain side.

Identification of Metabolites of Phytosterols in Rat Feces Using GC/MS

  • Song, Yun-Seon;Jin, Chang-bae;Park, Eun-Hee
    • Archives of Pharmacal Research
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    • v.23 no.6
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    • pp.599-604
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    • 2000
  • $\beta$-Sitosterol, campesterol and stigmasterol have been known to the phytosterols the most frequently found in plants. Metabolism of phytosterols was investigated using rat feces and liver microsomes. Feces were collected after phytosterols (a well characterized mixture of $\beta$-sitosterol 40%, campesterol 30% and dihydrobrasicasterol) were administered orally (0.5 ${g/kg$) to rats. Metabolites of phytosterols were identified using GC/MS. Three peaks were eluted at 12.47, 12.65, 12.87 min and had characteristic molecular ions m/z 428, 430, 432, respectively. Three fecal metabolites were identified as androstadienedione, androstenedione, and androstanedione. No metabolites could be detected in the rat liver microsomal reaction mixture. The results suggest that the metabolites of phytosterols in rat feces are formed by oxidation at 3- position, saturation at 5- and 6- position, and 17- side chain cleavage in the rat large intestine.

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Enzyme-Assisted Delignification of Several Pulps by Laccase from Botrytis Cinerea

  • Kim, Myung-Kil
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.34 no.2
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    • pp.67-72
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    • 2002
  • The two stage laccase-assisted delignification process led to significant lignin removal in the non-pressurized treatments. It is clearly shown that an alkaline extraction prior to the second laccase treatment significantly increased the overall delignification by ∼15%. This is in line with the contention that the residual lignin has undergone structural changes during the alkaline extraction, and the resulting modified structures are susceptible to the laccase oxidation. In phenolic hydroxyl group, the pre- methylated sample was very responsive to the delignification process. The phenolic hydroxyl groups could be increased during side chain cleavage catalyzed by laccase. This finding demonstrates that the delignification oi etherified structures is an important reaction in the delignificaton by laccase.