A dominant negative ADIPOQ mutation in a diabetic family with renal disease, hypoadiponectinemia, and hyperceramidemia

Figures

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  Fig. 1. Pedigree of the family enriched for diabetes and end-stage renal disease and identification of the ADIPOQ mutation. a The family pedigree, status of diabetes (shaded) and end-stage renal disease (ESRD; red outline), and the ADIPOQ mutation (p.Gly93GlufsTer73). Carriers of the wild-type ADIPOQ insertion (I) or mutated ADIPOQ deletion (D; red) are indicated. b The structure of the wild-type (top) and mutant (bottom) adiponectin protein. Wild-type adiponectin monomers consist of a 244 amino acid protein composed of four domains; an N-terminal signal sequence (19 amino acids), a variable region (23 amino acids), a collagenous domain (66 amino acids), and a C-terminal globular domain (136 amino acids). The 10-nucleotide deletion (CCCGAGGCTTT→C, indicated as ∆CCGAGGCTTT) at amino acid 93 creates a frameshift that truncates the adiponectin protein and generates a novel peptide that terminates 73 amino acids after this deletion. c The PHEVOR plot using Human Phenotype Ontology (HPO) terms and connections to Gene Ontology terms to prioritize potentially damaging alleles, using terms for a) kidney disease: HP:0000077 (Abnormality of the kidney), HP:0000112 (Nephropathy), and HP:0003774…

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  Fig. 2. Circulating adiponectin and ceramide levels and characterization of the ADIPOQ mutation. a The level of circulating adiponectin, measured by enzyme-linked immunosorbent assay (ELISA), in noncarriers (black, n = 7) and carriers (red, n = 8) of the ADIPOQ mutation (two-tailed t-test, α = 0.05, *p value ≤ 0.05, error bars SEM). b The intensity values of bands from western blotting for noncarrier (black, n = 2) and carrier (red, n = 2) patients. Carriers of the ADIPOQ mutation lack bands in FPLC fractions associated with high molecular weight (HMW) adiponectin complexes (error bars SEM). Representative western blots are shown and area under the curve was calculated for each peak, HMW, low molecular weight (LMW), and adiponectin trimers (2-way ANOVA with Fisher’s LSD correction, α = 0.05, p = 0.018, error bars are SEM). c The levels of circulating adiponectin, measured by ELISA, in noncarriers (black, n = 4) and carriers (red, n = 10) of the ADIPOQ mutation relative to C16.0 ceramide levels, measured by LC-MS/MS. d Western blot showing wild-type and mutant adiponectin produced in HEK293T cells. The upper blot represents cell lysates and the lower blot shows the two proteins excreted…