title: Structure of neuroblastoma suppressor of tumorigenicity 1 (NBL1): insights for the functional variability across bone morphogenetic protein (BMP) antagonists.
abstract: Bone morphogenetic proteins (BMPs) are antagonized through the action of numerous extracellular protein antagonists, including members from the differential screening-selected gene aberrative in neuroblastoma (DAN) family. In vivo, misregulation of the balance between BMP signaling and DAN inhibition can lead to numerous disease states, including cancer, kidney nephropathy, and pulmonary arterial hypertension. Despite this importance, very little information is available describing how DAN family proteins effectively inhibit BMP ligands. Furthermore, our understanding for how differences in individual DAN family members arise, including affinity and specificity, remains underdeveloped. Here, we present the structure of the founding member of the DAN family, neuroblastoma suppressor of tumorigenicity 1 (NBL1). Comparing NBL1 to the structure of protein related to Dan and Cerberus (PRDC), a more potent BMP antagonist within the DAN family, a number of differences were identified. Through a mutagenesis-based approach, we were able to correlate the BMP binding epitope in NBL1 with that in PRDC, where introduction of specific PRDC amino acids in NBL1 (A58F and S67Y) correlated with a gain-of-function inhibition toward BMP2 and BMP7, but not GDF5. Although NBL1(S67Y) was able to antagonize BMP7 as effectively as PRDC, NBL1(S67Y) was still 32-fold weaker than PRDC against BMP2. Taken together, this data suggests that alterations in the BMP binding epitope can partially account for differences in the potency of BMP inhibition within the DAN family.
authors.affiliation: Department of Molecular Genetics, Biochemistry, and Microbiology, The University of Cincinnati, Cincinnati, Ohio 45267 and.
authors.forename: Kristof
authors.initials: K
authors.lastname: Nolan
authors.affiliation: Department of Molecular Genetics, Biochemistry, and Microbiology, The University of Cincinnati, Cincinnati, Ohio 45267 and.
authors.forename: Chandramohan
authors.initials: C
authors.lastname: Kattamuri
authors.affiliation: Department of Molecular Genetics, Biochemistry, and Microbiology, The University of Cincinnati, Cincinnati, Ohio 45267 and.
authors.forename: David M
authors.initials: DM
authors.lastname: Luedeke
authors.affiliation: Department of Molecular Genetics, Biochemistry, and Microbiology, The University of Cincinnati, Cincinnati, Ohio 45267 and.
authors.forename: Elizabeth B
authors.initials: EB
authors.lastname: Angerman
authors.affiliation: Perinatal Institute, Cincinnati Children's Research Foundation and Department of Pediatrics, The University of Cincinnati, Cincinnati, Ohio 45229.
authors.forename: Scott A
authors.initials: SA
authors.lastname: Rankin
authors.affiliation: Perinatal Institute, Cincinnati Children's Research Foundation and Department of Pediatrics, The University of Cincinnati, Cincinnati, Ohio 45229.
authors.forename: Mariana L
authors.initials: ML
authors.lastname: Stevens
authors.affiliation: Perinatal Institute, Cincinnati Children's Research Foundation and Department of Pediatrics, The University of Cincinnati, Cincinnati, Ohio 45229.
authors.forename: Aaron M
authors.initials: AM
authors.lastname: Zorn
authors.affiliation: Department of Molecular Genetics, Biochemistry, and Microbiology, The University of Cincinnati, Cincinnati, Ohio 45267 and. Electronic address: tom.thompson@uc.edu.
authors.forename: Thomas B
authors.initials: TB
authors.lastname: Thompson
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics; D055419:Bone Morphogenetic Protein 7 / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000235:genetics; D016466:CHO Cells; D018797:Cell Cycle Proteins; D006224:Cricetinae; D003412:Cricetulus; D016207:Cytokines; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000737:chemistry* / Q000235:genetics; D016296:Mutagenesis; D020125:Mutation, Missense; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry* / Q000235:genetics; D013329:Structure-Activity Relationship
chemical_list: C525718:BMP2 protein, human; C525819:BMP7 protein, human; D055396:Bone Morphogenetic Protein 2; D055419:Bone Morphogenetic Protein 7; D018797:Cell Cycle Proteins; D016207:Cytokines; C495107:GREM2 protein, human; D036341:Intercellular Signaling Peptides and Proteins; C490087:NBL1 protein, human; D011506:Proteins
keywords: Bone Morphogenetic Protein (BMP); Differential Screening-aberrative in Neuroblastoma (DAN); Extracellular Antagonism; Mutagenesis; Neuroblastoma Suppressor of Tumorigenicity 1 (NBL1); Protein Related to Dan and Cerberus (PRDC); SOST; Structural Biology; Transforming Growth Factor β(TGF-B); X-ray Crystallography