Document Detail

High-level expression and purification of rat monoamine oxidase A (MAO A) in Pichia pastoris: comparison with human MAO A.
MedLine Citation:
PMID:  19883764     Owner:  NLM     Status:  MEDLINE    
The high-level heterologous expression in Pichia pastoris, purification and characterization of recombinant membrane-bound rat liver monoamine oxidase A (MAO A) are described. A 1-L culture of cells produces approximately 700 U of rat MAO A activity. The rat MAO A activity is found in outer mitochondrial membrane of the cell. Using a modification of the human MAO A purification procedure, approximately 200mg of recombinant rat MAO A is purified in a 43% yield and exhibits a molecular weight of approximately 60,000 kDa on SDS-PAGE. The purified enzyme contains a covalently bound FAD and forms a N(5) flavocyanine adduct on inhibition by clorgyline. Edman sequencing shows that the amino terminus of rat MAO A is blocked at an N-terminal threonyl residue. The purified rat enzyme exhibits a higher thermal stability than does purified human MAO A. Compared with human MAO A, rat MAO A oxidizes serotonin or kynuramine with twofold higher k(cat)/K(m) values, oxidizes phenethylamine with a 6.7-fold higher catalytic efficiency and benzylamine with a approximately 40-fold higher catalytic efficiency. Although approximately 90% identical in sequence to human MAO A, rat MAO A is a more efficient catalyst for amine neurotransmitter oxidation.
Jin Wang; Dale E Edmondson
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, N.I.H., Extramural     Date:  2009-10-31
Journal Detail:
Title:  Protein expression and purification     Volume:  70     ISSN:  1096-0279     ISO Abbreviation:  Protein Expr. Purif.     Publication Date:  2010 Apr 
Date Detail:
Created Date:  2010-02-23     Completed Date:  2010-05-20     Revised Date:  2014-09-16    
Medline Journal Info:
Nlm Unique ID:  9101496     Medline TA:  Protein Expr Purif     Country:  United States    
Other Details:
Languages:  eng     Pagination:  211-7     Citation Subset:  IM    
Copyright Information:
(c) 2009 Elsevier Inc. All rights reserved.
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MeSH Terms
Benzylamines / metabolism
Cloning, Molecular
Enzyme Stability
Monoamine Oxidase / biosynthesis*,  genetics,  isolation & purification,  metabolism
Phenethylamines / metabolism
Pichia / metabolism
Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
Substrate Specificity
Grant Support
Reg. No./Substance:
0/Benzylamines; 0/Phenethylamines; 327C7L2BXQ/phenethylamine; A1O31ROR09/benzylamine; EC Oxidase

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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