124-20-9Relevant articles and documents
Probing mammalian spermine oxidase enzyme-substrate complex through molecular modeling, site-directed mutagenesis and biochemical characterization
Tavladoraki, Paraskevi,Cervelli, Manuela,Antonangeli, Fabrizio,Minervini, Giovanni,Stano, Pasquale,Federico, Rodolfo,Mariottini, Paolo,Polticelli, Fabio
, p. 1115 - 1126 (2011)
Spermine oxidase (SMO) and acetylpolyamine oxidase (APAO) are FAD-dependent enzymes that are involved in the highly regulated pathways of polyamine biosynthesis and degradation. Polyamine content is strictly related to cell growth, and dysfunctions in polyamine metabolism have been linked with cancer. Specific inhibitors of SMO and APAO would allow analyzing the precise role of these enzymes in polyamine metabolism and related pathologies. However, none of the available polyamine oxidase inhibitors displays the desired characteristics of selective affinity and specificity. In addition, repeated efforts to obtain structural details at the atomic level on these two enzymes have all failed. In the present study, in an effort to better understand structure-function relationships, SMO enzyme-substrate complex has been probed through a combination of molecular modeling, site-directed mutagenesis and biochemical studies. Results obtained indicate that SMO binds spermine in a similar conformation as that observed in the yeast polyamine oxidase FMS1-spermine complex and demonstrate a major role for residues His82 and Lys367 in substrate binding and catalysis. In addition, the SMO enzyme-substrate complex highlights the presence of an active site pocket with highly polar characteristics, which may explain the different substrate specificity of SMO with respect to APAO and provide the basis for the design of specific inhibitors for SMO and APAO.
Mechanistic and structural analyses of the roles of active site residues in yeast polyamine oxidase Fms1: Characterization of the N195A and D94N enzymes
Adachi, Mariya S.,Taylor, Alexander B.,Hart, P. John,Fitzpatrick, Paul F.
, p. 8690 - 8697 (2012)
Flavoprotein Fms1 from Saccharomyces cerevisiae catalyzes the oxidation of spermine in the biosynthetic pathway for pantothenic acid. The same reaction is catalyzed by the mammalian polyamine and spermine oxidases. The active site of Fms1 contains three amino acid residues positioned to interact with the polyamine substrate, His67, Asn195, and Asp94. These three residues form a hydrogen-bonding triad with Asn195 being the central residue. Previous studies of the effects of mutating His67 are consistent with that residue being important both for interacting with the substrate and for maintaining the hydrogen bonds in the triad [Adachi, M. S., Taylor, A. B., Hart, P. J., and Fitzpatrick, P. F. (2012) Biochemistry 51, 4888-4897]. The N195A and D94N enzymes have now been characterized to evaluate their roles in catalysis. Both mutations primarily affect the reductive half-reaction. With N 1-acetylspermine as the substrate, the rate constant for flavin reduction decreases ~450-fold for both mutations; the effects with spermine as the substrate are smaller, 20-40-fold. The kcat/Kamine- and kcat-pH profiles with N1-acetylspermine are only slightly changed from the profiles for the wild-type enzyme, consistent with the pKa values arising from the amine substrate or product and not from active site residues. The structure of the N195A enzyme was determined at a resolution of 2.0 ?. The structure shows a molecule of tetraethylene glycol in the active site and establishes that the mutation has no effect on the protein structure. Overall, the results are consistent with the role of Asn195 and Asp94 being to properly position the polyamine substrate for oxidation.
A lysine conserved in the monoamine oxidase family is involved in oxidation of the reduced flavin in mouse polyamine oxidase
Henderson Pozzi, Michelle,Fitzpatrick, Paul F.
, p. 83 - 88 (2010)
Lysine 315 of mouse polyamine amine oxidase corresponds to a lysine residue that is conserved in the flavoprotein amine oxidases of the monoamine oxidase structural family. In several structures, this lysine residue forms a hydrogen bond to a water molecule that is hydrogen-bonded to the flavin N(5). Mutation of Lys315 in polyamine oxidase to methionine was previously shown to have no effect on the kinetics of the reductive half-reaction of the enzyme (M. Henderson Pozzi, V. Gawandi, P.F. Fitzpatrick, Biochemistry 48 (2009) 1508-1516). In contrast, the mutation does affect steps in the oxidative half-reaction. The kcat value is unaffected by the mutation; this kinetic parameter likely reflects product release. At pH 10, the kcat/Km value for oxygen is 25-fold lower in the mutant enzyme. The kcat/KO2 value is pH-dependent for the wild-type enzyme, decreasing below a pKa of 7.0, while this kinetic parameter for the mutant enzyme is pH-independent. This is consistent with the neutral form of Lys315 being required for more rapid flavin oxidation. The solvent isotope effect on the kcat/KO2 value increases from 1.4 in the wild-type enzyme to 1.9 in the mutant protein, and the solvent inventory changes from linear to bowed. The effects of the mutation can be explained by the lysine orienting the bridging water so that it can accept the proton from the flavin N(5) during flavin oxidation. In the mutant enzyme the lysine amine would be replaced by a water chain.
Preparation method of spermidine
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Paragraph 0032-0037, (2021/12/07)
The invention relates to the technical field of preparation of compounds, in particular to a preparation method of spermidine. The method comprises the following steps: S1, reacting 2-pyrrolidone and di-tert-butyl dicarbonate ester in an organic solvent by using 4-dimethylaminopyridine as a catalyst to obtain 1-(tert-butoxycarbonyl)-2-pyrrolidone; s2, dissolving the obtained 1-(tert-butoxycarbonyl)-2-pyrrolidone in a solvent, adding the solution into a 1, 3-propane diamine solution, carrying out stirring and reaction until the reaction is finished, and carrying out post-treatment to obtain [3-(3-amino-alanyl carbamoyl) propyl]-tert-butyl carbamate; s3, dissolving in a solvent, reducing by a reducing agent, and performing post-treatment to obtain [4-(3-amino-propylamino) butyl]-tert-butyl carbamate; and S4, dissolving in a solvent, dropwise adding hydrochloric acid for reaction to remove a protecting group, and after the reaction, further treating to obtain the spermidine. The raw materials such as pyrrolidone, 1, 3-propane diamine and the like are common chemical raw materials, the cost is relatively low, the reaction conditions are relatively mild, and the potential safety hazard in the reaction is relatively low.
Method for preparing spermidine
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Paragraph 0057; 0072-0074, (2019/01/14)
The invention belongs to the field of chemical synthesis and particularly relates to a method for preparing spermidine. The method comprises the steps that the spermidine is prepared through the mainprocess steps of reduction, amino protection and the like after reaction of amino-1-propanol and butyrolactone serving as raw materials. The method has the advantages of being simple in operation, high in product quality, high in yield and the like.