Situs inversus totalis(SIT)is a rare homozygous recessive disease caused by the mutation in PKD1L1,which is required for normal interaction with PKD2 and leads to different complications such as respiratory disorders,...Situs inversus totalis(SIT)is a rare homozygous recessive disease caused by the mutation in PKD1L1,which is required for normal interaction with PKD2 and leads to different complications such as respiratory disorders,brain disorders and even obesity.The present study was designed to find out the mutational effect on the binding of PKD2 with mutated PKD1L1,which leads to SIT.The three-dimensional(3D)structure of wild type and mutated PKD1L1 was predicted with>90%confidence using different online tools.The different online tools that were employed were SWISS-MODEL,Phyre2(normal&intensive)and i-TASSER.To compute the physiochemical properties of PKD1L1(wild&mutated)and PKD2 in silico approaches were employed using the ExPASy ProtParam tool.Physicochemical properties such as molecular weight,isoelectric point,the total number of negatively and positively charged residues,extinction coefficient,half-life,instability and aliphatic index,grand average of hydropathicity,and amino acid percentage were calculated.A lot of variability was observed in these parameters among PKD1L1 and PKD2,which accounted for diversification in their functional properties.The theoretical pI points showed that PKD1L1(whole)is more basic with 6.64 pI compared to its first chain TOPO_DOM(amino acids from 1–1748)has a pI of 5.62 which means it is basic while PKD2 have the lowest pI point of 5.34.Docking was performed using the PatchDock and ClusPro online tools.展开更多
Objective: To study the expression of PKD2 gene in human kidney and other tissues. Methods: The expression of PKD2 was detected by reverse transcription PCR(RT-PCR) and in situ hybridization(ISH) . The results of ISH ...Objective: To study the expression of PKD2 gene in human kidney and other tissues. Methods: The expression of PKD2 was detected by reverse transcription PCR(RT-PCR) and in situ hybridization(ISH) . The results of ISH were analyzed by micromegakargooytes. Results: Distribution of pkd-2 in normal adult kidney was stronger in proximal convoluted tubule, Henle's loop ascending branch, distal convoluted tubule and cortical collecting ducts, and inferior signal were observed in fetal kidney. Negative was seen in ADPKD 2 kidney. Conclusion: Down-regulation of PKD2 gene expression in kidney may take effect on the occurrence and development of ADPKD2.展开更多
Objective:Autosomal dominant polycystic kidney disease(ADPKD)is mainly caused by the pathogenic mutation of PKD1 or PKD2 gene and usually affects bilateral kidneys.Synonymous mutations are generally assumed to be neut...Objective:Autosomal dominant polycystic kidney disease(ADPKD)is mainly caused by the pathogenic mutation of PKD1 or PKD2 gene and usually affects bilateral kidneys.Synonymous mutations are generally assumed to be neutral as they do not alter amino acids.Herein,we described an extremely rare ADPKD child caused by a heterozygous synonymous mutation of PKD2 gene accompanied by massive proteinuria and congenital solitary kidney.Methods:Clinical characteristics of the patients were summarized.Whole-exome sequencing was performed to screen the disease-causing gene mutation,and reverse transcription polymerase chain reaction(RT-PCR)and Sanger sequencing were applied to analyze the impact of the identified mutation on gene transcription and splicing.Results:Polycystic changes were found in the solitary kidney of a girl initially presented with nephrotic-range proteinuria.Thereafter her mother and 2 other family members were diagnosed to be ADPKD.Whole-exome sequencing of the proband identified a heterozygous synonymous mutation(c.1716G>A,p.Lys572=)located in the splicing site of exon 7 in PKD2 gene,which was co-segregated with the PKD phenotype in the family.RT-PCR and direct sequencing of amplified products revealed that this heterozygous synonymous mutation led to exon7 skipping in PKD2 gene.Conclusion:We reported an extremely rare child case of ADPKD2 in combination with solitary kidney and nephrotic-range proteinuria,and firstly confirmed the pathogenicity of a heterozygous synonymous mutation(c.1716G>A)in PKD2 gene.The results indicate that synonymous mutations should not be excluded from disease-causing if they are located in splicing site of an exon.展开更多
文摘Situs inversus totalis(SIT)is a rare homozygous recessive disease caused by the mutation in PKD1L1,which is required for normal interaction with PKD2 and leads to different complications such as respiratory disorders,brain disorders and even obesity.The present study was designed to find out the mutational effect on the binding of PKD2 with mutated PKD1L1,which leads to SIT.The three-dimensional(3D)structure of wild type and mutated PKD1L1 was predicted with>90%confidence using different online tools.The different online tools that were employed were SWISS-MODEL,Phyre2(normal&intensive)and i-TASSER.To compute the physiochemical properties of PKD1L1(wild&mutated)and PKD2 in silico approaches were employed using the ExPASy ProtParam tool.Physicochemical properties such as molecular weight,isoelectric point,the total number of negatively and positively charged residues,extinction coefficient,half-life,instability and aliphatic index,grand average of hydropathicity,and amino acid percentage were calculated.A lot of variability was observed in these parameters among PKD1L1 and PKD2,which accounted for diversification in their functional properties.The theoretical pI points showed that PKD1L1(whole)is more basic with 6.64 pI compared to its first chain TOPO_DOM(amino acids from 1–1748)has a pI of 5.62 which means it is basic while PKD2 have the lowest pI point of 5.34.Docking was performed using the PatchDock and ClusPro online tools.
基金Supported by the 10th Five Year Plan Program for Major Sci-tech Foundation(No. 2002AAgZ3130)National INatural Science Founda- tion of China (No. 30170901, No.30271523)The Hundred Leading Scientists Program of the Public Health Sector of Shanghai (No. 97BR047)Major Basic Research Foundation of Shanghai Science and Technology Committee (No.02JC14029)
文摘Objective: To study the expression of PKD2 gene in human kidney and other tissues. Methods: The expression of PKD2 was detected by reverse transcription PCR(RT-PCR) and in situ hybridization(ISH) . The results of ISH were analyzed by micromegakargooytes. Results: Distribution of pkd-2 in normal adult kidney was stronger in proximal convoluted tubule, Henle's loop ascending branch, distal convoluted tubule and cortical collecting ducts, and inferior signal were observed in fetal kidney. Negative was seen in ADPKD 2 kidney. Conclusion: Down-regulation of PKD2 gene expression in kidney may take effect on the occurrence and development of ADPKD2.
基金supported by the National Natural Science Foundation of China(No.81873596).
文摘Objective:Autosomal dominant polycystic kidney disease(ADPKD)is mainly caused by the pathogenic mutation of PKD1 or PKD2 gene and usually affects bilateral kidneys.Synonymous mutations are generally assumed to be neutral as they do not alter amino acids.Herein,we described an extremely rare ADPKD child caused by a heterozygous synonymous mutation of PKD2 gene accompanied by massive proteinuria and congenital solitary kidney.Methods:Clinical characteristics of the patients were summarized.Whole-exome sequencing was performed to screen the disease-causing gene mutation,and reverse transcription polymerase chain reaction(RT-PCR)and Sanger sequencing were applied to analyze the impact of the identified mutation on gene transcription and splicing.Results:Polycystic changes were found in the solitary kidney of a girl initially presented with nephrotic-range proteinuria.Thereafter her mother and 2 other family members were diagnosed to be ADPKD.Whole-exome sequencing of the proband identified a heterozygous synonymous mutation(c.1716G>A,p.Lys572=)located in the splicing site of exon 7 in PKD2 gene,which was co-segregated with the PKD phenotype in the family.RT-PCR and direct sequencing of amplified products revealed that this heterozygous synonymous mutation led to exon7 skipping in PKD2 gene.Conclusion:We reported an extremely rare child case of ADPKD2 in combination with solitary kidney and nephrotic-range proteinuria,and firstly confirmed the pathogenicity of a heterozygous synonymous mutation(c.1716G>A)in PKD2 gene.The results indicate that synonymous mutations should not be excluded from disease-causing if they are located in splicing site of an exon.