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Polymorphisms of SLC22A9 (hOAT7) in Korean Females with Osteoporosis.

Ahn SK, Suh CK, Cha SH - Korean J. Physiol. Pharmacol. (2015)

Bottom Line: The net decrease of estrogen, especially in post-menopausal women induces rapid bone loss.The G767A, T1277C and C1487T SNPs result in an amino acid substitution, from synonymous vs nonsynonymous substitution arginine to glutamine (R256Q), phenylalanine to serine (F426S) and proline to leucine (P496L), respectively.The Km values and Vmax of the wild type, R256Q, P496L and F426S were 8.84, 8.87, 9.83 and 12.74 µM, and 1.97, 1.96, 2.06 and 1.55 pmol/oocyte/h, respectively.

View Article: PubMed Central - PubMed

Affiliation: Department of Tropical Medicine and Parasitology, College of Medicine, Inha University, Incheon 400-712, Korea.

ABSTRACT
Among solute carrier proteins, the organic anion transporters (OATs) play an important role for the elimination or reabsorption of endogenous and exogenous negatively charged anionic compounds. Among OATs, SLC22A9 (hOAT7) transports estrone sulfate with high affinity. The net decrease of estrogen, especially in post-menopausal women induces rapid bone loss. The present study was performed to search the SNP within exon regions of SLC22A9 in Korean females with osteoporosis. Fifty healthy controls and 50 osteoporosis patients were screened for the genetic polymorphism in the coding region of SLC22A9 using GC-clamped PCR and denaturing gradient gel electrophoresis (DGGE). Six SNPs were found on the SLC22A9 gene from Korean women with/without osteoporosis. The SNPs were located as follows: two SNPs in the osteoporosis group (A645G and T1277C), three SNPs in the control group (G1449T, C1467T and C1487T) and one SNP in both the osteoporosis and control groups (G767A). The G767A, T1277C and C1487T SNPs result in an amino acid substitution, from synonymous vs nonsynonymous substitution arginine to glutamine (R256Q), phenylalanine to serine (F426S) and proline to leucine (P496L), respectively. The Km values and Vmax of the wild type, R256Q, P496L and F426S were 8.84, 8.87, 9.83 and 12.74 µM, and 1.97, 1.96, 2.06 and 1.55 pmol/oocyte/h, respectively. The present study demonstrates that the SLC22A9 variant F426S is causing inter-individual variation that is leading to the differences in transport of the steroid sulfate conjugate (estrone sulfate) and, therefore this could be used as a marker for certain disease including osteoporosis.

No MeSH data available.


Related in: MedlinePlus

The typical band patterns of agarose or acrylamide gel electrophoresis. (A) Normal individual genomic DNA amplified using DGGE primer sets for the respective mutation found in exons by duplex PCR. (B) The duplex PCR products from the same genomic DNAs were loaded on a polyacrylamide gel. (C) The aberrant band patterns and sequencing electropherograms of observed SLC22A9 SNPs in osteoporosis and normal subjects. The synonymous A645G, G1449T and C1467T were found in exon 3 and 9, respectively. The G767A, T1277C and C1487T were found in exon 4, 7 and 9, respectively.
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Figure 2: The typical band patterns of agarose or acrylamide gel electrophoresis. (A) Normal individual genomic DNA amplified using DGGE primer sets for the respective mutation found in exons by duplex PCR. (B) The duplex PCR products from the same genomic DNAs were loaded on a polyacrylamide gel. (C) The aberrant band patterns and sequencing electropherograms of observed SLC22A9 SNPs in osteoporosis and normal subjects. The synonymous A645G, G1449T and C1467T were found in exon 3 and 9, respectively. The G767A, T1277C and C1487T were found in exon 4, 7 and 9, respectively.

Mentions: The DGGE method was used in order to determine the SNP. The PCR conditions were tested over several annealing temperatures and cycle numbers using normal human chromosomal DNA. The typical patterns of each PCR products with synonymous and non-synonymous mutations found using GC-clamping primer sets separated on 1% agarose gel are presented in Fig. 2A. The representative results of DGGE separated on acrylamide gel using several control samples are shown in Fig. 2B. The results of DGGE showed a single band in the SNP found in exons.


Polymorphisms of SLC22A9 (hOAT7) in Korean Females with Osteoporosis.

Ahn SK, Suh CK, Cha SH - Korean J. Physiol. Pharmacol. (2015)

The typical band patterns of agarose or acrylamide gel electrophoresis. (A) Normal individual genomic DNA amplified using DGGE primer sets for the respective mutation found in exons by duplex PCR. (B) The duplex PCR products from the same genomic DNAs were loaded on a polyacrylamide gel. (C) The aberrant band patterns and sequencing electropherograms of observed SLC22A9 SNPs in osteoporosis and normal subjects. The synonymous A645G, G1449T and C1467T were found in exon 3 and 9, respectively. The G767A, T1277C and C1487T were found in exon 4, 7 and 9, respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4499643&req=5

Figure 2: The typical band patterns of agarose or acrylamide gel electrophoresis. (A) Normal individual genomic DNA amplified using DGGE primer sets for the respective mutation found in exons by duplex PCR. (B) The duplex PCR products from the same genomic DNAs were loaded on a polyacrylamide gel. (C) The aberrant band patterns and sequencing electropherograms of observed SLC22A9 SNPs in osteoporosis and normal subjects. The synonymous A645G, G1449T and C1467T were found in exon 3 and 9, respectively. The G767A, T1277C and C1487T were found in exon 4, 7 and 9, respectively.
Mentions: The DGGE method was used in order to determine the SNP. The PCR conditions were tested over several annealing temperatures and cycle numbers using normal human chromosomal DNA. The typical patterns of each PCR products with synonymous and non-synonymous mutations found using GC-clamping primer sets separated on 1% agarose gel are presented in Fig. 2A. The representative results of DGGE separated on acrylamide gel using several control samples are shown in Fig. 2B. The results of DGGE showed a single band in the SNP found in exons.

Bottom Line: The net decrease of estrogen, especially in post-menopausal women induces rapid bone loss.The G767A, T1277C and C1487T SNPs result in an amino acid substitution, from synonymous vs nonsynonymous substitution arginine to glutamine (R256Q), phenylalanine to serine (F426S) and proline to leucine (P496L), respectively.The Km values and Vmax of the wild type, R256Q, P496L and F426S were 8.84, 8.87, 9.83 and 12.74 µM, and 1.97, 1.96, 2.06 and 1.55 pmol/oocyte/h, respectively.

View Article: PubMed Central - PubMed

Affiliation: Department of Tropical Medicine and Parasitology, College of Medicine, Inha University, Incheon 400-712, Korea.

ABSTRACT
Among solute carrier proteins, the organic anion transporters (OATs) play an important role for the elimination or reabsorption of endogenous and exogenous negatively charged anionic compounds. Among OATs, SLC22A9 (hOAT7) transports estrone sulfate with high affinity. The net decrease of estrogen, especially in post-menopausal women induces rapid bone loss. The present study was performed to search the SNP within exon regions of SLC22A9 in Korean females with osteoporosis. Fifty healthy controls and 50 osteoporosis patients were screened for the genetic polymorphism in the coding region of SLC22A9 using GC-clamped PCR and denaturing gradient gel electrophoresis (DGGE). Six SNPs were found on the SLC22A9 gene from Korean women with/without osteoporosis. The SNPs were located as follows: two SNPs in the osteoporosis group (A645G and T1277C), three SNPs in the control group (G1449T, C1467T and C1487T) and one SNP in both the osteoporosis and control groups (G767A). The G767A, T1277C and C1487T SNPs result in an amino acid substitution, from synonymous vs nonsynonymous substitution arginine to glutamine (R256Q), phenylalanine to serine (F426S) and proline to leucine (P496L), respectively. The Km values and Vmax of the wild type, R256Q, P496L and F426S were 8.84, 8.87, 9.83 and 12.74 µM, and 1.97, 1.96, 2.06 and 1.55 pmol/oocyte/h, respectively. The present study demonstrates that the SLC22A9 variant F426S is causing inter-individual variation that is leading to the differences in transport of the steroid sulfate conjugate (estrone sulfate) and, therefore this could be used as a marker for certain disease including osteoporosis.

No MeSH data available.


Related in: MedlinePlus