• Title/Summary/Keyword: Small protein

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Effects of Korean Ginseng Saponin Fraction on the Biosynthesis of Spermidine and Spermine from Bat Prostate and Testis

  • 조윤식;조영대
    • Journal of Ginseng Research
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    • v.22 no.4
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    • pp.316-323
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    • 1998
  • To study e(feces of Korean Binseng (Parfax ginseff C. A. hfeyer) total saponin fraction on spermidine and spermine metabolism in rat reproductive systems, we administrated the saponin fractation to rats for 2 years. Then, we determined the activities of S-adenosylmethionine decarboxylase (SAMDC), the quantitation of the enzyme protein and the amounts of spermidine and spermine contents In prostate and testis. In young sexually immature stage, administration of Korean ginseng saponin fraction showed no effect on SAMDC activities. The stimulatory effect on the activities of SAMDC gradually increased and reached maximal activities in test groups of prostate and testis at sexually mature stage. The amounts of SAMDC protein in test groups were paralleled by the changes of SAMDC activities in test groups, indicating that all of the increased activity occurring in administration of ginseng saponin fraction was not due to the activation of SAMDC activity but to the Increase in enzyme protein. However, the spermidine and spermine contents of test groups showed small increase in compared to that of control groups. From these results, we suggest that administration of ginseng saponin fraction alter the spermidine and spermine metabolism in sexually mature and aged reproductive systems in rats.

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Post-Translational Modification of Proteins in Toxicological Research: Focus on Lysine Acylation

  • Lee, Sangkyu
    • Toxicological Research
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    • v.29 no.2
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    • pp.81-86
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    • 2013
  • Toxicoproteomics integrates the proteomic knowledge into toxicology by enabling protein quantification in biofluids and tissues, thus taking toxicological research to the next level. Post-translational modification (PTM) alters the three-dimensional (3D) structure of proteins by covalently binding small molecules to them and therefore represents a major protein function diversification mechanism. Because of the crucial roles PTM plays in biological systems, the identification of novel PTMs and study of the role of PTMs are gaining much attention in proteomics research. Of the 300 known PTMs, protein acylation, including lysine formylation, acetylation, propionylation, butyrylation, malonylation, succinylation, and crotonylation, regulates the crucial functions of many eukaryotic proteins involved in cellular metabolism, cell cycle, aging, growth, angiogenesis, and cancer. Here, I reviewed recent studies regarding novel types of lysine acylation, their biological functions, and their applicationsin toxicoproteomics research.

Evaluation of proteomic strategies for analyzing ubiquitinated proteins

  • Peng, Jun Min
    • BMB Reports
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    • v.41 no.3
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    • pp.177-183
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    • 2008
  • Ubiquitin is an essential, highly-conserved small regulatory protein in eukaryotic cells. It covalently modifies a wide variety of targeted proteins in the forms of monomer and polymers, altering the conformation and binding properties of the proteins and thus regulating proteasomal delivery, protein activities and localization. Mass spectrometry has emerged as an indispensable tool for in-depth characterization of protein ubiquitination. Ubiquitinated proteins in cell lysates are usually enriched by affinity chromatography and subsequently analyzed by mass spectrometry for identification and quantification. Ubiquitin-conjugated amino acid residues can be determined by unique mass shift caused by the modification. Moreover, the complex structure of polyubiquitin chains on substrates can be dissected by bottom-up and middle-down mass spectrometric approaches, revealing potential novel functions of polyubiquitin linkages. Here I review the advances and caveats of these strategies, emphasizing caution in the validation of ubiquitinated proteins and in the interpretation of raw data.

Multifactorial Regulation of G Protein-Coupled Receptor Endocytosis

  • Zhang, Xiaohan;Kim, Kyeong-Man
    • Biomolecules & Therapeutics
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    • v.25 no.1
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    • pp.26-43
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    • 2017
  • Endocytosis is a process by which cells absorb extracellular materials via the inward budding of vesicles formed from the plasma membrane. Receptor-mediated endocytosis is a highly selective process where receptors with specific binding sites for extracellular molecules internalize via vesicles. G protein-coupled receptors (GPCRs) are the largest single family of plasma-membrane receptors with more than 1000 family members. But the molecular mechanisms involved in the regulation of GPCRs are believed to be highly conserved. For example, receptor phosphorylation in collaboration with ${\beta}$-arrestins plays major roles in desensitization and endocytosis of most GPCRs. Nevertheless, a number of subsequent studies showed that GPCR regulation, such as that by endocytosis, occurs through various pathways with a multitude of cellular components and processes. This review focused on i) functional interactions between homologous and heterologous pathways, ii) methodologies applied for determining receptor endocytosis, iii) experimental tools to determine specific endocytic routes, iv) roles of small guanosine triphosphate-binding proteins in GPCR endocytosis, and v) role of post-translational modification of the receptors in endocytosis.

Backbone NMR Assignments of WW2 domain from human AIP4

  • Seo, Min-Duk
    • Journal of the Korean Magnetic Resonance Society
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    • v.24 no.2
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    • pp.38-42
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    • 2020
  • WW domains are small protein modules consisting of three-stranded antiparallel β-sheet, and involved in the protein-protein interaction for various biological systems. We overexpressed and purified WW2 domain from human AIP4/Itch (a member of Nedd4 family) using a pH/temperature dependent cleavage system. The backbone assignments of WW2 domain were completed, and secondary structure was predicted. Furthermore, backbone flexibility of WW2 domain was determined by 1H-15N heteronuclear NOE and amide hydrogen exchange experiments. The structural information would contribute to the structural determination of WW2 domain as well as the interaction study of WW2 domain with various binding partners.

Modulation of Rit Activation by the Alpha Subunit of Go

  • Yang, Chul-Min;Ghil, Sung-Ho
    • Biomedical Science Letters
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    • v.15 no.4
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    • pp.327-333
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    • 2009
  • Heterotrimeric GTP binding proteins, G-proteins, mediate signal transduction generated by neurotransmitters and hormones. Among G-proteins, Go proteins are the most abundant in brain and classified as a member of Gi family. Ras-like protein in all tissues (Rit), one of the small GTPases, is a member of a Ras superfamily and identified as an important regulator of neuronal differentiation and cell transformation. Recently, we have reported that Rit functioned as a candidate downstream effector for alpha subunit of Go proteins ($Go{\alpha}$) and regulated neurite outgrowth triggered by $Go{\alpha}$ activation. In this study, we showed that the GTPase domain of $Go{\alpha}$ contributed to the direct interaction with Rit. We also demonstrated that $Go{\alpha}$ could lead to an increase of Rit activity suggesting that Rit play a role as a downstream effector of $Go{\alpha}$.

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A QUADRATIC APPROXIMATION FOR PROTEIN SEQUENCE TO STRUCTURE MAPPING

  • Oh, Se-Young;Yun, Jae-Heon;Chung, Sei-Young
    • Journal of applied mathematics & informatics
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    • v.12 no.1_2
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    • pp.155-164
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    • 2003
  • A method is proposed to predict the distances between given residue pairs (between C$\sub$${\alpha}$/ atoms) of a protein using a sequence to structure mapping by indefinite quadratic approximation. The prediction technique requires a data fitting in three dimensional space with coordinates of the residues of known structured proteins and leads to a numerical ref resentation of 20 amino acids by minimizing a large least norm iteratively. These approximations are used in distance prediction for given residue pairs. Some computational experience on a test set of small proteins from Brookhaven Protein Data Bank are given.

Isolation and Characterization of a Cryptic Plasmid, pMBLR00, from Leuconostoc mesenteroides subsp. mesenteroides KCTC 3733

  • Chae, Han Seung;Lee, Jeong Min;Lee, Ju-Hoon;Lee, Pyung Cheon
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.837-842
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    • 2013
  • A cryptic plasmid, pMBLR00, from Leuconostoc mesenteroides subsp. mesenteroides KCTC 3733 was isolated, characterized, and used for the construction of a cloning vector to engineer Leuconostoc species. pMBLR00 is a rolling circle replication plasmid, containing 3,370 base pairs. Sequence analysis revealed that pMBLR00 has 3 open reading frames: Cop (copy number control protein), Rep (replication protein), and Mob (mobilization protein). pMBLR00 replicates by rolling circle replication, which was confirmed by the presence of a conserved double-stranded origin and single-stranded DNA intermediates. An Escherichia coli-Leuconostoc shuttle vector, pMBLR02, was constructed and was able to replicate in Leuconostoc citreum 95. pMBLR02 could be a useful genetic tool for metabolic engineering and the genetic study of Leuconostoc species.

BINDING OF LEAF NUCLEAR PROTEIN EXTRACTS TO LIGHT-RESPONSIVIE ELEMENTS OF cab PROMOTERS OF Arabidopsis thaliana

  • Lee, Hwa-Hyung;Park, Hee-Jin
    • Journal of Photoscience
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    • v.3 no.2
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    • pp.85-90
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    • 1996
  • The binding ability of leaf nuclear extracts to the lighbresponsive elements (LREs) of cab promoters of Arabidopsis thaliana has been investigated. The cab promoters were fragmented with restr ction endonucleases into LRE that were identified by Mitra et al. [Plant Mol. Biol. 12, 169179 ( 1989)] and other small fragments. After end labeling with Klenow fragment, the fragments were assayed for binding with the leaf nuclear proteins that were prepared by solubilizing the purified nuclei with 0.5 M ammonium sulfate. The binding ability was assayed by mobility shift assay. To perform successful mobility shift assay, several factors affecting the interaction of protein with DNA were optimized before performing the assay. The LREs had several retardation bands. However, the other promoter fragments from the transcription start site to the far upstream region of the promoters had also retardation bands. No particular relationships could be found between the retardation band distributions and the loci of LRE. It is likely that the light-regulation of cab gene expression may be controlled by the multiple interactions of the regulatory protein factors with DNA motifs.

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Improvement of crop traits using auxin binding protein gene abp57 (옥신 호르몬 결합단백질 ABP57 유전자를 이용한 작물의 형질개선)

  • Kim, Dong-Hern;Lee, Keun-Pyo
    • Journal of Plant Biotechnology
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    • v.38 no.2
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    • pp.137-142
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    • 2011
  • Auxin is a group of small natural and synthetic molecules having diverse regulatory functions in plant growth and development. In this review, two auxin binding proteins identified by biochemical experiments to measure their auxin binding activities and biochemical functions are described. ABP1, a 22 kDa auxin binding protein, shows strong auxin binding affinity and possibly plays an important role in plant development, although its biochemical function are still unclear. ABP57, a 57 kDa soluble protein from rice shoots, has both of IAA binding activity and the plasma membrane proton pump activation. Although it is yet to be accomplished, the improvement of agronomic traits using auxin binding proteins is worth to be considered, since auxin is known to be related to such a diverse crop traits.