5K25

Crystal structure of human phosphatase PRL-2 in complex with the ADP-bound Bateman domain of human magnesium transporter CNNM3

Summary for 5K25

• Entry DOI: 10.2210/pdb5k25/pdb
• Related: 5K22 5K23 5K24
• Descriptor: Protein tyrosine phosphatase type IVA 2, Metal transporter CNNM3, ADENOSINE-5'-DIPHOSPHATE, ... (4 entities in total)
• Functional Keywords: complex, phosphatase, magnesium transporter, protein binding, hydrolase, hydrolase-transporter protein-protein binding complex, hydrolase/transporter protein/protein binding
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 2
• Total formula weight: 40116.69

Authors

Gulerez, I.,Kozlov, G.,Gehring, K. (deposition date: 2016-05-18, release date: 2016-10-12, Last modification date: 2024-10-30)

Primary citation

Zhang, H.,Kozlov, G.,Li, X.,Wu, H.,Gulerez, I.,Gehring, K.
PRL3 phosphatase active site is required for binding the putative magnesium transporter CNNM3.
Sci Rep, 7:48-48, 2017

PubMed Abstract: The phosphatases of regenerating liver (PRLs) are involved in tumorigenesis and metastatic cancer yet their cellular function remains unclear. Recent reports have shown PRL phosphatases bind tightly to the CNNM family of membrane proteins to regulate magnesium efflux. Here, we characterize the interactions between the CBS-pair (Bateman) domain of CNNM3 and either PRL2 or PRL3 using X-ray crystallography, isothermal titration calorimetry, and activity assays. We report four new crystal structures of PRL proteins bound to the CNNM3 CBS-pair domain that reveal the effects of cysteine disulphide formation and nucleotide binding on complex formation. We use comprehensive mutagenesis of the PRL3 catalytic site to quantify the importance of different PRL amino acids, including cysteine 104, leucine 108, and arginine 110, for CNNM binding and phosphatase activity. We show the PRL3 R138E mutant is selectively deficient in CNNM3 binding with the potential to distinguish between the downstream effects of phosphatase and CNNM-binding activities in vivo. Through a novel activity assay, we show that PRL3 has magnesium-sensitive phosphatase activity with ATP and other nucleotides. Our results identify a strong correlation between phosphatase activity and CNNM binding and support the contention that PRL function as pseudophosphatases regulated by chemical modifications of their catalytic cysteine.

PubMed: 28246390
DOI: 10.1038/s41598-017-00147-2

Experimental method

X-RAY DIFFRACTION (3.05 Å)