Linked Life Data

12779324

Source:http://linkedlifedata.com/resource/pubmed/id/12779324

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
22
pubmed:dateCreated
2003-6-3
pubmed:abstractText
Patatin is a nonspecific lipid acyl hydrolase that accounts for approximately 40% of the total soluble protein in mature potato tubers, and it has potent insecticidal activity against the corn rootworm. We determined the X-ray crystal structure of a His-tagged variant of an isozyme of patatin, Pat17, to 2.2 A resolution, employing SeMet multiwavelength anomalous dispersion (MAD) phasing methods. The patatin crystal structure has three molecules in the asymmetric unit, an R-factor of 22.0%, and an R(free) of 27.2% (for 10% of the data not included in the refinement) and includes 498 water molecules. The structure notably revealed that patatin has a Ser-Asp catalytic dyad and an active site like that of human cytosolic phospholipase A(2) (cPLA(2)) [Dessen, A., et al. (1999) Cell 97, 349-360]. In addition, patatin has a folding topology related to that of the catalytic domain of cPLA(2) and unlike the canonical alpha/beta-hydrolase fold. The structure confirms our site-directed mutagenesis and bioactivity data that initially suggested patatin possessed a Ser-Asp catalytic dyad. Alanine-scanning mutagenesis revealed that Ser77 and Asp215 were critical for both esterase and bioactivity, consistent with prior work implicating a Ser residue [Strickland, J. H., et al. (1995) Plant Physiol. 109, 667-674] and a Ser-Asp dyad [Hirschberg, H. J. H. B., et al. (2001) Eur. J. Biochem. 268, 5037-5044] in patatin's catalytic activity. The crystal structure aids the understanding of other structure-function relationships in patatin. Patatin does not display interfacial activation, a hallmark feature of lipases, and this is likely due to the fact that it lacks a flexible lid that can shield the active site.
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Jun
pubmed:issn
0006-2960
pubmed:author
pubmed:issnType
Print
pubmed:day
10
pubmed:volume
42
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
6696-708
pubmed:dateRevised
2006-11-15
pubmed:meshHeading
pubmed-meshheading:12779324-Alanine, pubmed-meshheading:12779324-Amino Acid Substitution, pubmed-meshheading:12779324-Animals, pubmed-meshheading:12779324-Aspartic Acid, pubmed-meshheading:12779324-Beetles, pubmed-meshheading:12779324-Carboxylic Ester Hydrolases, pubmed-meshheading:12779324-Catalytic Domain, pubmed-meshheading:12779324-Cloning, Molecular, pubmed-meshheading:12779324-Crystallography, X-Ray, pubmed-meshheading:12779324-Humans, pubmed-meshheading:12779324-Isoenzymes, pubmed-meshheading:12779324-Larva, pubmed-meshheading:12779324-Models, Molecular, pubmed-meshheading:12779324-Mutagenesis, Site-Directed, pubmed-meshheading:12779324-Phospholipases A, pubmed-meshheading:12779324-Pichia, pubmed-meshheading:12779324-Plant Proteins, pubmed-meshheading:12779324-Protein Structure, Secondary, pubmed-meshheading:12779324-Serine
pubmed:year
2003
pubmed:articleTitle
The crystal structure, mutagenesis, and activity studies reveal that patatin is a lipid acyl hydrolase with a Ser-Asp catalytic dyad.
pubmed:affiliation
Monsanto Company, Chesterfield, Missouri 63017-1732, USA. timothy.j.rydel@monsanto.com
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, Non-P.H.S., Research Support, Non-U.S. Gov't