title: Different conformational dynamics of PDZ1 and PDZ2 in full-length EBP50 analyzed by hydrogen/deuterium exchange mass spectrometry.
abstract: Ezrin-radixin-moesin-binding protein 50 (EBP50) is a scaffolding protein expressed in polarized epithelial cells in various organs, including the liver, kidney, and small intestine, in which it regulates the trafficking and targeting cellular proteins. EBP50 contains two postsynaptic density-95/disk-large/ZO-1 homology (PDZ) domains (e.g., PDZ1 and PDZ2) and an ezrin/radixin/moesin-binding (EB) domain. PDZ domains are one of the major scaffolding domains regulating protein-protein interactions with critical biological roles in cell polarity, migration, proliferation, recognition, and cell-cell interaction. PDZ1 and PDZ2 in EBP50 have different ligand selectivity, although several high-resolution structural studies of isolated PDZ1 and PDZ2 showed similar structures. We studied the conformations of full-length EBP50 and isolated PDZ1 and PDZ2 using hydrogen/deuterium exchange mass spectrometry (HDX-MS). The deuterium uptake profiles of isolated PDZ1 and PDZ2 were similar to those of full-length EBP50. Interestingly, PDZ1 was more dynamic than PDZ2, and these PDZ domains underwent different conformational changes upon ligand binding. These results might explain the differences in ligand-selectivity between PDZ1 and PDZ2.
authors: a School of Pharmacy, Sungkyunkwan University, 2066 Seobu-ro, Jangan-gu, Suwon, 440746, Republic of Korea.
authors: Ji Young
authors: JY
authors: Park
authors: a School of Pharmacy, Sungkyunkwan University, 2066 Seobu-ro, Jangan-gu, Suwon, 440746, Republic of Korea.
authors: Nguyen Minh
authors: NM
authors: Duc
authors: a School of Pharmacy, Sungkyunkwan University, 2066 Seobu-ro, Jangan-gu, Suwon, 440746, Republic of Korea.
authors: Dong Kyun
authors: DK
authors: Kim
authors: a School of Pharmacy, Sungkyunkwan University, 2066 Seobu-ro, Jangan-gu, Suwon, 440746, Republic of Korea.
authors: Su Youn
authors: SY
authors: Lee
authors: b Department of medicine, University of California at San Diego, La Jolla, CA 92093, USA.
authors: Sheng
authors: S
authors: Li
authors: c College of Pharmacy & Department of Molecular Science and Technology, Ajou University, Suwon 443749, Republic of Korea.
authors: Min-Duk
authors: MD
authors: Seo
authors: b Department of medicine, University of California at San Diego, La Jolla, CA 92093, USA.
authors: Virgil L
authors: VL
authors: Woods
authors: a School of Pharmacy, Sungkyunkwan University, 2066 Seobu-ro, Jangan-gu, Suwon, 440746, Republic of Korea.
authors: Ka Young
authors: KY
authors: Chung
mesh_terms: D000595:Amino Acid Sequence; D003903:Deuterium; D006801:Humans; D006859:Hydrogen; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D054731:PDZ Domains; D010750:Phosphoproteins / Q000737:chemistry*; D011487:Protein Conformation; D017923:Sodium-Hydrogen Exchangers / Q000737:chemistry*
chemical_list: D010750:Phosphoproteins; D017923:Sodium-Hydrogen Exchangers; C093376:sodium-hydrogen exchanger regulatory factor; D006859:Hydrogen; D003903:Deuterium
keywords: EBP50; PDZ domain; domaine PDZ; hydrogen/deuterium exchange mass spectrometry; structure; échange hydrogène/deutérium couplé à la spectrométrie de masse