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
Matrix metalloproteinas-9 (MMP-9) is a glycosylated endopeptidase, and hence its processing between the endoplasmic reticulum (ER), Golgi and trans-Golgi (TGN) network remains under a strict control of factors that af...Matrix metalloproteinas-9 (MMP-9) is a glycosylated endopeptidase, and hence its processing between the endoplasmic reticulum (ER), Golgi and trans-Golgi (TGN) network remains under a strict control of factors that affect the microtubule (MT) stabilization, and the recruitment and activation of coat and cargo proteins, including ADP-ribosylation factors (Arfs) and protein kinase D (PKD). Here, we report on the factors implicated in the regulation of MMP-9 secretion by salivary gland acinar cells in response to P. gingivalis LPS, and the effect of hormone, ghrelin. We show that the LPS-elicited induction in MMP-9 secretion is associated with the increase in α-tubulin acetylation and the enhancement in MT stabilization, while the modulatory effect of ghrelin is reflected in a decrease in α-tubulin acetylation. Further, the effect of the LPS occurs in concert with up-regulation in Arf-guanine nucleotide exchange factor (GEF)-mediated Arf1 activation and the TGN recruitment of PKD2, while ghrelin exerts the modulatory effect on Arf-GEF activation. Moreover, we reveal that the LPS-induced up-regulation in MMP-9 secretion is reflected in a marked increase in PKCδ-mediated PKD2 phosphorylation on Ser, while the modulatory effect of ghrelin is manifested by the SFK-PTKs-dependent phosphorylation of PKD2 on Tyr. The findings demonstrate that MT stabilization along with Arf-GEF-mediated Arf1/PKD2 activation play a major role in P. gingivalis LPS-induced up-regulation in salivary gland acinar cell MMP-9 secretion, and point the modulatory mode of action by ghrelin.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
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.
文摘Matrix metalloproteinas-9 (MMP-9) is a glycosylated endopeptidase, and hence its processing between the endoplasmic reticulum (ER), Golgi and trans-Golgi (TGN) network remains under a strict control of factors that affect the microtubule (MT) stabilization, and the recruitment and activation of coat and cargo proteins, including ADP-ribosylation factors (Arfs) and protein kinase D (PKD). Here, we report on the factors implicated in the regulation of MMP-9 secretion by salivary gland acinar cells in response to P. gingivalis LPS, and the effect of hormone, ghrelin. We show that the LPS-elicited induction in MMP-9 secretion is associated with the increase in α-tubulin acetylation and the enhancement in MT stabilization, while the modulatory effect of ghrelin is reflected in a decrease in α-tubulin acetylation. Further, the effect of the LPS occurs in concert with up-regulation in Arf-guanine nucleotide exchange factor (GEF)-mediated Arf1 activation and the TGN recruitment of PKD2, while ghrelin exerts the modulatory effect on Arf-GEF activation. Moreover, we reveal that the LPS-induced up-regulation in MMP-9 secretion is reflected in a marked increase in PKCδ-mediated PKD2 phosphorylation on Ser, while the modulatory effect of ghrelin is manifested by the SFK-PTKs-dependent phosphorylation of PKD2 on Tyr. The findings demonstrate that MT stabilization along with Arf-GEF-mediated Arf1/PKD2 activation play a major role in P. gingivalis LPS-induced up-regulation in salivary gland acinar cell MMP-9 secretion, and point the modulatory mode of action by ghrelin.
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.
