Characterization of a Compound Heterozygous SLC2A9 Mutation That Causes Hypouricemia
Abstract
:1. Introduction
2. Methods
2.1. Study Participants
2.2. DNA Preparation and Whole-Exome Sequencing
2.3. WES Variant Filtering Analysis
2.4. Direct Sanger Sequencing
2.5. In Silico Analysis of Novel Missense Variants and Molecular Dynamics
2.5.1. In Silico Prediction
2.5.2. Molecular Dynamics
2.5.3. Molecular/Functional Studies
Generation of SLC2A9b Expression Vectors
Site-Directed Mutagenesis for the Met126Val Mutant
In Vitro Transcription
SLC2A9b Expression in X. laevis Oocytes
Western Blot Analysis
Uric Acid Uptake Assay
Confocal Microscopy
3. Results
3.1. Demographics
3.1.1. Identification of Novel Variants in SLC2A9b by Whole-Exome Sequencing
3.1.2. In Silico and Molecular Dynamics Prediction of SLC2A9b
3.1.3. Molecular Dynamics Prediction of SLC2A9b and Its Affinity for Uric Acid
3.2. Molecular Analysis
3.2.1. SLC2A9b-p.Met126Val Expression Analysis in X. laevis Oocytes
3.2.2. Urate Transport Activity of SLC2A9b-p.Met126Val in Xenopus Oocytes
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Characteristics | Unexplained Group | SLC2A9 Compound Heterozygote |
---|---|---|
n = 6 | n = 1 | |
Age (years) | 43 ± 12 | 40 |
BMI † (kg/m2) | 25.1 ± 2.9 | 23.8 |
Waist circumference, cm | 81 ± 5 | 72 |
Blood pressure, mmHg | ||
Systolic | 110 ± 3 | 124 |
Diastolic | 71 ± 12 | 64 |
Smoking status | ||
Never a smoker, no. (%) | 1 (16.67) | 1 (100) |
Ever a smoker, no. (%) | 5 (83.33) | 0 (0) |
Alcohol consumption | ||
Never a drinker, no. (%) | 2 (33.33) | 1 (100) |
Ever a drinker, no. (%) | 4 (66.67) | 0 (0) |
Uric acid, mg/dL | 0.78 ± 0.52 | 0.80 |
Total cholesterol, mg/dL | 214 ± 34 | 174 |
Triglycerides, mg/dL | 169 ± 69 | 178 |
Fasting glucose, mg/dL | 86 ± 14 | 92 |
LDL cholesterol, mg/dL | 116 ± 22 | 90.4 |
HDL cholesterol, mg/dL | 64 ± 18 | 48 |
Creatinine, mg/dL | 0.80 ± 0.25 | 0.70 |
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Yoon, J.; Cachau, R.; David, V.A.; Thompson, M.; Jung, W.; Jee, S.-H.; Daar, I.O.; Winkler, C.A.; Cho, S.-K. Characterization of a Compound Heterozygous SLC2A9 Mutation That Causes Hypouricemia. Biomedicines 2021, 9, 1172. https://doi.org/10.3390/biomedicines9091172
Yoon J, Cachau R, David VA, Thompson M, Jung W, Jee S-H, Daar IO, Winkler CA, Cho S-K. Characterization of a Compound Heterozygous SLC2A9 Mutation That Causes Hypouricemia. Biomedicines. 2021; 9(9):1172. https://doi.org/10.3390/biomedicines9091172
Chicago/Turabian StyleYoon, Jaeho, Raul Cachau, Victor A. David, Mary Thompson, Wooram Jung, Sun-Ha Jee, Ira O. Daar, Cheryl A. Winkler, and Sung-Kweon Cho. 2021. "Characterization of a Compound Heterozygous SLC2A9 Mutation That Causes Hypouricemia" Biomedicines 9, no. 9: 1172. https://doi.org/10.3390/biomedicines9091172