TY - JOUR
T1 - Crystal structure of phospholipase A1 from Streptomyces albidoflavus NA297
AU - Murayama, Kazutaka
AU - Kano, Kota
AU - Matsumoto, Yusaku
AU - Sugimori, Daisuke
N1 - Funding Information:
We thank Associate Professor Chiaki Ogino (Kobe University, Japan) for providing the expression vector. The synchrotron radiation experiments were performed at the Photon Factory (Tsukuba, Japan). We would like to thank the beamline staff at BL-1A of the Photon Factory for assistance during data collection. We also acknowledge the support of the Biomedical Research Core of Tohoku University, Graduate School of Medicine . This work was partly supported by the Targeted Proteins Research Program from the Ministry of Education, Culture, Sports, Science and Technology, Japan .
PY - 2013/5
Y1 - 2013/5
N2 - The metal-independent lipase from Streptomyces albidoflavus NA297 (SaPLA1) is a phospholipase A1 as it preferentially hydrolyzes the sn-1 acyl ester in glycerophospholipids, yielding a fatty acid and 2-acyl-lysophospholipid. The molecular mechanism underlying the substrate binding by SaPLA1 is currently unknown. In this study, the crystal structure of SaPLA1 was determined at 1.75å resolutions by molecular replacement. A structural similarity search indicated the highest structural similarity to an esterase from Streptomyces scabies, followed by GDSL family enzymes. The SaPLA1 active site is composed of a Ser-His dyad (Ser11 and His218), whereby stabilization of the imidazole is provided by the main-chain carbonyl oxygen of Ser216, a common variation of the catalytic triad in many serine hydrolases, where this carbonyl maintains the orientation of the active site histidine residue. The hydrophobic pocket and cleft for lipid binding are adjacent to the active site, and are approximately 13-15å deep and 14-16å long. A partial polyethylene glycol structure was found in this hydrophobic pocket.
AB - The metal-independent lipase from Streptomyces albidoflavus NA297 (SaPLA1) is a phospholipase A1 as it preferentially hydrolyzes the sn-1 acyl ester in glycerophospholipids, yielding a fatty acid and 2-acyl-lysophospholipid. The molecular mechanism underlying the substrate binding by SaPLA1 is currently unknown. In this study, the crystal structure of SaPLA1 was determined at 1.75å resolutions by molecular replacement. A structural similarity search indicated the highest structural similarity to an esterase from Streptomyces scabies, followed by GDSL family enzymes. The SaPLA1 active site is composed of a Ser-His dyad (Ser11 and His218), whereby stabilization of the imidazole is provided by the main-chain carbonyl oxygen of Ser216, a common variation of the catalytic triad in many serine hydrolases, where this carbonyl maintains the orientation of the active site histidine residue. The hydrophobic pocket and cleft for lipid binding are adjacent to the active site, and are approximately 13-15å deep and 14-16å long. A partial polyethylene glycol structure was found in this hydrophobic pocket.
KW - Catalytic dyad
KW - Hydrophobic pocket
KW - Metal-independent lipase
KW - Phospholipase A
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U2 - 10.1016/j.jsb.2013.02.003
DO - 10.1016/j.jsb.2013.02.003
M3 - Article
C2 - 23416196
AN - SCOPUS:84876741452
SN - 1047-8477
VL - 182
SP - 192
EP - 196
JO - Journal of Structural Biology
JF - Journal of Structural Biology
IS - 2
ER -