---
reference_id: PMID:12779324
title: "The crystal structure, mutagenesis, and activity studies reveal that patatin is a lipid acyl hydrolase with a Ser-Asp catalytic dyad."
authors:
- Rydel TJ
- Williams JM
- Krieger E
- Moshiri F
- Stallings WC
- Brown SM
- Pershing JC
- Purcell JP
- Alibhai MF
journal: Biochemistry
year: '2003'
doi: 10.1021/bi027156r
content_type: abstract_only
full_text_attempted: true
---

# The crystal structure, mutagenesis, and activity studies reveal that patatin is a lipid acyl hydrolase with a Ser-Asp catalytic dyad.
**Authors:** Rydel TJ, Williams JM, Krieger E, Moshiri F, Stallings WC, Brown SM, Pershing JC, Purcell JP, Alibhai MF
**Journal:** Biochemistry (2003)
**DOI:** [10.1021/bi027156r](https://doi.org/10.1021/bi027156r)

## Content

1. Biochemistry. 2003 Jun 10;42(22):6696-708. doi: 10.1021/bi027156r.

The crystal structure, mutagenesis, and activity studies reveal that patatin is 
a lipid acyl hydrolase with a Ser-Asp catalytic dyad.

Rydel TJ(1), Williams JM, Krieger E, Moshiri F, Stallings WC, Brown SM, Pershing 
JC, Purcell JP, Alibhai MF.

Author information:
(1)Monsanto Company, Chesterfield, Missouri 63017-1732, USA. 
timothy.j.rydel@monsanto.com

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.

DOI: 10.1021/bi027156r
PMID: 12779324 [Indexed for MEDLINE]