Structural basis of substrate discrimination and integrin binding by autotaxin.
Autotaxin (ATX, also known as ectonucleotide pyrophosphatase/phosphodiesterase-2, ENPP2) is a secreted lysophospholipase D that generates the lipid mediator lysophosphatidic acid (LPA), a mitogen and chemoattractant for many cell types. ATX-LPA signaling is involved in various pathologies including tumor progression and inflammation. However, the molecular basis of substrate recognition and catalysis by ATX and the mechanism by which it interacts with target cells are unclear. Here, we present the crystal structure of ATX, alone and in complex with a small-molecule inhibitor. We have identified a hydrophobic lipid-binding pocket and mapped key residues for catalysis and selection between nucleotide and phospholipid substrates. We have shown that ATX interacts with cell-surface integrins through its N-terminal somatomedin B-like domains, using an atypical mechanism. Our results define determinants of substrate discrimination by the ENPP family, suggest how ATX promotes localized LPA signaling and suggest new approaches for targeting ATX with small-molecule therapeutic agents.
Division of Biochemistry, The Netherlands Cancer Institute, Amsterdam, The Netherlands.
Jens
J
Hausmann
Satwik
S
Kamtekar
Evangelos
E
Christodoulou
Jacqueline E
JE
Day
Tao
T
Wu
Zachary
Z
Fulkerson
Harald M H G
HM
Albers
Laurens A
LA
van Meeteren
Anna J S
AJ
Houben
Leonie
L
van Zeijl
Silvia
S
Jansen
Maria
M
Andries
Troii
T
Hall
Lyle E
LE
Pegg
Timothy E
TE
Benson
Mobien
M
Kasiem
Karl
K
Harlos
Craig W Vander
CW
Kooi
Susan S
SS
Smyth
Huib
H
Ovaa
Mathieu
M
Bollen
Andrew J
AJ
Morris
Wouter H
WH
Moolenaar
Anastassis
A
Perrakis
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D020134:Catalytic Domain; D002460:Cell Line; D018360:Crystallography, X-Ray; D006801:Humans; D016023:Integrins / Q000378:metabolism*; D008246:Lysophospholipids / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D010727:Phosphoric Diester Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011755:Pyrophosphatases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D013379:Substrate Specificity; D000099189:Lysophospholipase D
D016023:Integrins; D008246:Lysophospholipids; D010727:Phosphoric Diester Hydrolases; D011755:Pyrophosphatases; D000099189:Lysophospholipase D; C032881:lysophosphatidic acid
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