• Proceedings of the Korean Society for Bioinformatics Conference
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• 2003.10a
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• pp.5-5
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• 2003

The purple acid phosphatases comprise a family of binuclear metal-containing enzymes. The metal centre contains one ferric ion and one divalent metal ion. Spectroscopic studies of the monomeric, ${\sim}$36 kDa mammalian purple acid phosphatases reveal the presence of an Fe(III)Fe(II) centre in which the metals are weakly antiferromagnetically coupled, whereas the dimeric, ${\sim}$110 000 kDa plant enzymes contain either Fe(III)Zn(II) or Fe(III)Mn(II). The three dimensional structures of the red kidney bean and pig enzymes show very similar arrangements of the metal ligands but some significant differences beyond the immediate vicinity of the metals. In addition to the catalytic domain, the plant enzyme contains a second domain of unknown function. A search of sequence databases was undertaken using a sequence pattern which includes the conserved metal-binding residues in the plant and animal enzymes. The search revealed the presence in plants of a 'mammalian-type' low molecular weight purple acid phosphatase, a high molecular weight form in some fungi, and a homologue in some bacteria. The catalytic mechanism of the enzyme has been investigated with a view to understanding the marked difference in specificity between the Fe-Mn sweet potato enzyme, which exhibits highly efficient catalysis towards both activated and unactivated phosphate esters, and other PAPs, which hydrolyse only activated esters. Comparison of the active site structures of the enzymes reveal some interesting differences between them which may account for the difference. The implications fur understanding the physiological functions of the enzymes will be discussed.

• Applied Chemistry for Engineering
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• v.9 no.6
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• pp.803-810
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• 1998

We have studied the catalytic combustion of soot particulate over perovskite-type oxides prepared by malic acid method. The catalysts were modified to enhance the activity by substitution of metal into A or B site of perovskite oxide. In addition, the reaction conditions, such as temperature, $O_2$ concentration, space velocity have been studied. The effect of $SO_2$ pretreatment and water introduced into reactants were also examined. In the $LaCoO_3$ catalyst, the partial substitution of alkali metals into A site enhanced the catalytic activity in the combustion of soot particulate and the activity was shown in the order : Cs>K>Na; In the $La_{0.6}Cs_{0.4}CoO_3 $; catalyst, the substitution of Fe or Mn showed no effect on the ignition temperature. The ignition temperature decreased with increasing $O_2$ concentration and contact time. The introduction of water into reactants feed decreased the ignition temperature and the pretreatment of $SO_2$ showed no effect on the catalytic activity.

• Proceedings of the Korean Society of Applied Pharmacology
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• 1996.11a
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• pp.135-139
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• 1996

$p70^{s6k}$ lies on a $p21^{ras}$-independent signalling pathway and plays an important role in mitogenesis. Activation is associated with phosphorylation at multiple sites, four of which lie in an autoinhibitory region. The immunosuppressant rapamycin induces $p70^{s6k}$ inactivation through dephosphorylation of a second set of mitogen-induced sites. Here we identify these sites as $T_{229}$, $T_{389}$, and $S_{404}$. $T_{229}$ resides in the "T loop" of the catalytic domain, an essential phosphorylation site in other kinases. However, $p70^{s6k}$ inactivation by rapamycin most closely parallels $T_{389}$ dephosphorylation. Mutation of $T_{389}$ to alanine ablates kinase activity, whereas mutation to glutamic acid confers constitutive kinase activity and rapamycin resistance. indicating an essential role for phosphorylation at this site. $T_{389}$ resides in an unusual hydrophobic motif, not previously noted, between the catalytic and autoinhibitory domains. The importance of this site, and surrounding motif, is emphasized by its conservation in other kinases including homologues of $p70^{s6k}$ derived from such distantly related organisms as yeast and plant.

• BMB Reports
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• v.41 no.1
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• pp.48-54
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• 2008

PM0188 is a newly identified sialyltransferase from P. multocida which transfers sialic acid from cytidine 5'-monophosphonuraminic acid (CMP-NeuAc) to an acceptor sugar. Although sialyltransferases are involved in important biological functions like cell-cell recognition, cell differentiation and receptor-ligand interactions, little is known about their catalytic mechanism. Here, we report the X-ray crystal structures of PM0188 in the presence of an acceptor sugar and a donor sugar analogue, revealing the precise mechanism of sialic acid transfer. Site-directed mutagenesis, kinetic assays, and structural analysis show that Asp141, His311, Glu338, Ser355 and Ser356 are important catalytic residues; Asp141 is especially crucial as it acts as a general base. These complex structures provide insights into the mechanism of sialyltransferases and the structure-based design of specific inhibitors.

• Applied Chemistry for Engineering
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• v.10 no.3
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• pp.407-414
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• 1999

We have studied the reduction of NO by CO over perovskite-type oxides prepared by malic and method. The catalysts were modified to enhance the activity by substitution of metal into A or B site of perovskite oxides. In the $LaCoO_3$ type catalyst, the partial substitution of Sr into A site enhanced the catalytic activity on the conversion of NO at less than $350^{\circ}C$. In the $La_{0.6}Sr_{0.4}Co_{1-x}Fe_xO_3$ catalyst, the partial substitution of Fe or Mn into B site enhanced the conversion of NO, but excess amount of Fe decreased the conversion of NO. In addition, $La_{0.6}Sr_{0.4}Co_{0.8}Fe_{0.2}O_3$ mixed with $SnO_2$ or $MnO_2$ showed the synergy effect on the reduction of NO. The introduction of water into reactants feed decreased the catalytic activity but the deactivation was shown to be reversible. The introduction of $SO_2$ into reactants feed also decreased the catalytic activity.

• BMB Reports
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• v.38 no.2
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• pp.248-252
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• 2005

Dihydrolipoamide dehydrogenase (E3) catalyzes the reoxidation of dihydrolipoyl moiety of the acyltransferase components of three $\alpha$-keto acid dehydrogenase complexes and of the hydrogen-carrier protein of the glycine cleavage system. His-457 of Pseudomonas putida E3 is suggested to interact with the hydroxyl group of Tyr-18 of the other subunit and with Glu-446, a component in the last helical structure. To examine the importance of the suggested interactions in human E3 function, the corresponding residue of human E3, Asn-473, was substituted to Leu using site-directed mutagenesis. The E3 mutant was expressed in Escherichia coli and highly purified using an affinity column. Its E3 activity was decreased about 37-fold, indicating that Asn-473 residue was important to the efficient catalytic function of human E3. Its slightly altered spectroscopic properties implied that small conformational changes could occur in the E3 mutant.

• BMB Reports
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• v.30 no.2
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• pp.113-118
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• 1997

Aspartase from Hafnia alvei was inactivated by N-ethylmaleimide (NEM) and 5,5' -Dithiobis-(2-znitrobenzoic acid) (DTNB) following pseudo-first order kinetics. Their apparent reaction orders were 0.83 and 0.50 for NEM and DTNB modifications, respectively, indicating that inactivation was due to a sulfhydryl group in the active site of aspartase and participation of the sulfhydryl group in an essential step in the catalytic reaction. When aspartase was modified by DTNB, the enzyme activity was restored by dithiothreitol treatment, indicating that cysteine residuetsl islarel possibly at or near the active site. The pH-dependence of the inactivation rate by NEM suggested that an amino acid residue having pK value of 8.3 was involved in the inactivation. When aspartase was incubated with NEM and L-aspartate together, L-aspartate markedly protected the enzyme from inactivation by NEM, but the other reagents used did not.

• BMB Reports
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• v.39 no.2
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• pp.223-227
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• 2006

Dihydrolipoamide dehydrogenase (E3) belongs to the pyridine nucleotide-disulfide oxidoreductase family including glutathione reductase and thioredoxin reductase. It catalyzes the reoxidation of dihydrolipoyl moiety of the acyltransferase components of three $\alpha$-keto acid dehydrogenase complexes and of the hydrogen-carrier protein of the glycine cleavage system. Isoleucine-51 of human E3, located near the active disulfide center Cys residues, is highly conserved in most E3s from several sources. To examine the importance of this highly conserved Ile-51 in human E3 function, it was substituted with Ala using site-directed mutagenesis. The mutant was expressed in Escherichia coli and highly purified using an affinity column. Its E3 activity was decreased about 100-fold, indicating that the conservation of the Ile-51 residue in human E3 was very important to the efficient catalytic function of the enzyme. Its altered spectroscopic properties implied that conformational changes could occur in the mutant.

• Journal of Microbiology and Biotechnology
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• v.26 no.5
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• pp.837-845
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• 2016

A novel endodextranase isolated from Paenibacillus sp. was found to produce isomaltotetraose and small amounts of cycloisomaltooligosaccharides with a degree of polymerization of 7-14 from dextran. To determine the active site, the enzyme was modified with 1-ethyl-3-[3-(dimethylamino)-propyl]-carbodiimide (EDC) and α-epoxyalkyl α-glucosides (EAGs), an affinity labeling reagent. The inactivation followed pseudo first-order kinetics. Kinetic analysis and chemical modification using EDC and EAGs indicated that carboxyl groups are essential for the enzymatic activity. Three Asp and one Glu residues were identified as candidate catalytic amino acids, since these residues are completely conserved across the GH family of 66 enzymes. Replacement of Asp189, Asp340, or Glu412 completely abolished the enzyme activity, indicating that these residues are essential for catalytic activity.

• Journal of Life Science
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• v.21 no.3
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• pp.473-479
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• 2011

Adenylyl cyclase (AC) catalyzes the formation of cyclic AMP (cAMP) from ATP. The cAMP produced by AC serves as a secondary messenger in a variety of signal transduction pathways, and controls various cellular functions in many organisms. ACs can be grouped into six classes based on their primary amino acid sequences. Eukaryotes and mycobacteria contain only members of class III AC. The catalytic cyclase domains of class III AC are active as dimers: mammalian ACs, which are composed of a single polypeptide with two catalytic cyclase domains, form the active site as a result of intramolecular dimerization of the catalytic cyclase domains. In contrast, mycobacterial ACs function as homodimers, since their polypeptides contain a single catalytic cyclase domain. Six amino acids are required for the catalytic activity of class III AC - two aspartate residues, a lysine-aspartate pair and an arginine-asparagine pair. 16 ACs belonging to the class III were identified in Mycobacterium tuberculosis H37Rv, and their characteristics are reviewed.