Extended Data Fig. 4: Isolation and functional validation of LPP heterooligomers.
From: Structural basis for the catalytic mechanism of human lipid phosphate phosphatases
a, Sequence alignment of human LPP1, LPP2, and LPP3 highlights the conservation of essential residues involved in tetramerization and the coordination of PAE, PAI, and PIP2 lipids. These residues are indicated by blue squares, purple circles, red circles, and red triangles, respectively. The C1, C2, and C3 loops are marked with brown, deep blue-green, and marine boxes, respectively. b, Structural overlay of the LPP1 structure with the Alphafold2-predicted models of LPP2 and LPP3. The Alphafold2-predicted models were downloaded from the UniProt database (https://www.uniprot.org/). c, Immunoblotting analysis demonstrated successful isolation of LPP1-LPP2, LPP1-LPP3, and LPP2-LPP3 heterooligomers. The representative blots shown here are from three independent experiments. Please refer to the Methods section for details. d, SEC analysis of these heterooligomers suggests that they exhibit similar oligomeric states. e, Functional assays confirmed that these LPP heterooligomers retain the ability to dephosphorylate LPA, PA, and S1P. In this experiment, the enzymatic systems were incubated at 37 °C for 20 min prior to quantification using the malachite green-ammonium molybdate method. Data are presented as the means ± s.d. of three independent experiments, each with two technical repeats (n=6).
