In this study, we report that the insertion of a pseudoproline dipeptide for the solid-phase peptide synthesis of wild-type Phospholamban protein (WT-PLB) has two important advantages. First, it disrupts the formation...In this study, we report that the insertion of a pseudoproline dipeptide for the solid-phase peptide synthesis of wild-type Phospholamban protein (WT-PLB) has two important advantages. First, it disrupts the formation of different secondary structures, which is responsible for poor couplings during the preparation of highly aggregated sequences. Second, it enhances the purities and solubility of crude products leading to easier HPLC purification.展开更多
In this study, we report the microwave assisted synthesis of Phospholamban protein (WT-PLB) as a new gold standard in solid phase peptide synthesis. Microwave energy offers benefits for both the coupling and deprotect...In this study, we report the microwave assisted synthesis of Phospholamban protein (WT-PLB) as a new gold standard in solid phase peptide synthesis. Microwave energy offers benefits for both the coupling and deprotection reactions during peptide synthesis. The use of microwave energy for both the coupling and deprotection steps makes the microwave peptide synthesizers the most versatile and powerful systems available. It produces high yield and fast synthesis when compared to conventional peptide synthesizers.展开更多
Objective To study the effect of phospholamban antisense RNA (asPLB) on sarcoplasmic reticulum Ca2+-ATPase activity and cardiac function in rats with diabetes mellitus (DM) mediated by recombinant adeno-associated vir...Objective To study the effect of phospholamban antisense RNA (asPLB) on sarcoplasmic reticulum Ca2+-ATPase activity and cardiac function in rats with diabetes mellitus (DM) mediated by recombinant adeno-associated virus (rAAV) vector. Methods Six weeks after the induction of DM by streptozotocin injected intraperitoneally, the rats were divided into three groups, namely: DM-rAAV-asPLB group, DM-saline group and DM group (control group). The rats in the DM-rAAV-asPLB group were intramyocardially injected with rAAV-asPLB, the rats in the DM-saline group were injected with saline, and those in the control group did not receive any treatment. Six weeks after gene transfer, the expressions of PLB protein and PLB phosphorylation were detected by Western-blot, while the activity of sarcoplasmic reticulum (SR) Ca2+-ATPase and left ventricular function were measured. Results The PLB protein expression level was significantly higher whereas the PLB phosphorylation, SR Ca2+-ATPase activity and left ventricular function were significantly lower in the DM-saline group than in the control group. No significant difference was found in PLB protein expression level, PLB phosphorylation or SR Ca2+-ATPase activity between the DM-rAAV-asPLB group and the control group. The left ventricular function in the DM-rAAV-asPLB group was poorer than in the control group and was better than in the DM-saline group. Conclusion rAAV-asPLB can down-regulate PLB protein expression and up-regulate PLB phosphorylation and SR Ca2+ -ATPase activity, thus contributing to the improvement of in vivo left ventricular function.展开更多
This study aimed to determine the prevalence and clinical features of Arrhythmogenic Right Ventricular Cardiomyopathy(ARVC)caused by pathogenic mutations in the Phospholamban(PLN)gene.The study included 170 patients w...This study aimed to determine the prevalence and clinical features of Arrhythmogenic Right Ventricular Cardiomyopathy(ARVC)caused by pathogenic mutations in the Phospholamban(PLN)gene.The study included 170 patients who had a confrmed diagnosis of ARVC and underwent PLN genetic screening using next-generation sequencing.The fndings of this study provide valuable insights into the association between PLN mutations and ARVC,which can aid in the development of more efective diagnostic and treatment strategies for ARVC patients.Out of the patients evaluated,six had a rare pathogenic mutation in PLN with the same p.R14del variant.Family screening revealed that heterozygous carriers of p.R14del exhibited a defnite ARVC phenotype.In clinical studies,individuals with the p.R14del mutation experienced a similar rate of malignant arrhythmia events as those with classic desmosome mutations.After adjusting for covariates,individuals with PLN mutations had a two point one seven times greater likelihood of experiencing transplant-related risks compared to those who did not possess PLN mutations(95%CI 1.08–6.82,p=0.035).The accumulation of left ventricular fat and fbers is a pathological marker for ARVC patients with p.R14del mutations.In a cohort of 170 Chinese ARVC patients,three point fve percent of probands had the PLN pathogenic variant(p.R14del)and all were female.Our data shows that PLN-related ARVC patients are at high risk for ventricular arrhythmias and heart failure,which requires clinical diferentiation from classic ARVC.Furthermore,carrying the p.R14del mutation can be an independent prognostic risk factor in ARVC patients.展开更多
Phospholamban is an important protein with responsibility for regulating the activity of the sarcoplasmic reticulum Ca2+ pump through reversible phosphorylation.And its three-dimensional structure in living cell has b...Phospholamban is an important protein with responsibility for regulating the activity of the sarcoplasmic reticulum Ca2+ pump through reversible phosphorylation.And its three-dimensional structure in living cell has been a focus of attention.In the current case, we summarized the investigations on phospholamban structure, and on this base, employed long time-scale molecular dy-namics simulations to study its structure systematically.The first 22 residues from one chain of phospholamban in bellflower structure determined by NMR experiments, together with its phosphorylation at position 16 and mutation at position 9 were picked up as three different systems.By molecular dynamics simulations of 10 ns in the explicit solution surroundings, it was found that the 3–15 residues of the original structure retained their helix structures, while the phosphorylation and mutation had less probability to form helix structures.These structural changes might result in inhibition decrease to the sarcoplasmic reticulum Ca2+ pump, which is in accordance with previous experimental results.展开更多
目的:研究扩心方对阿霉素诱导扩张型心肌病模型大鼠心功能、心肌损伤的保护作用,探讨扩心方作用机制。方法:50只Wistar大鼠腹腔注射阿霉素6周建立DCM模型,开始造模4周后随机分为模型组,扩心方高、中、低剂量组,卡托普利组(每组各10只),...目的:研究扩心方对阿霉素诱导扩张型心肌病模型大鼠心功能、心肌损伤的保护作用,探讨扩心方作用机制。方法:50只Wistar大鼠腹腔注射阿霉素6周建立DCM模型,开始造模4周后随机分为模型组,扩心方高、中、低剂量组,卡托普利组(每组各10只),连续灌胃给药四周,另设10只正常对照。正常组和模型组以生理盐水灌胃。治疗结束后,对各组大鼠进行心脏超声检查,病理形态学检测,以及用反转录-聚合酶链反应和蛋白质免疫印迹法分别检测心肌组织中SERCA2α、PLB的m RNA和蛋白含量。结果:与正常对照组相比,模型组心功能减弱,出现心肌病理损害,SERCA2α与PLB表达异常;与模型组相比,扩心方组可以改善DCM大鼠的心功能、心肌病理学形态,同时,提高SERCA2αm RNA和蛋白表达、抑制PLB m RNA和蛋白表达。结论:扩心方能提高DCM大鼠的左心室射血分数和缩短率,减轻心肌损伤,改善心功能,同时调控DCM大鼠SERCA2α、PLB的表达。提示扩心方改善DCM大鼠心功能的作用机制,可能与纠正心肌肌浆网钙调控相关蛋白-SERCA2α、PLB的异常表达相关。展开更多
文摘In this study, we report that the insertion of a pseudoproline dipeptide for the solid-phase peptide synthesis of wild-type Phospholamban protein (WT-PLB) has two important advantages. First, it disrupts the formation of different secondary structures, which is responsible for poor couplings during the preparation of highly aggregated sequences. Second, it enhances the purities and solubility of crude products leading to easier HPLC purification.
文摘In this study, we report the microwave assisted synthesis of Phospholamban protein (WT-PLB) as a new gold standard in solid phase peptide synthesis. Microwave energy offers benefits for both the coupling and deprotection reactions during peptide synthesis. The use of microwave energy for both the coupling and deprotection steps makes the microwave peptide synthesizers the most versatile and powerful systems available. It produces high yield and fast synthesis when compared to conventional peptide synthesizers.
基金supported by Scientific Research Foundation of College of Medicine,Zhejiang UniversityFoundation of Zhejiang Science and Technology Bureau(No.2007B071)+1 种基金Foundation of Zhejiang Educational Committee(No.Y200908338)Foundation of Zhejiang Traditional Chinese Medicine Committee(No.2011B503103)
文摘Objective To study the effect of phospholamban antisense RNA (asPLB) on sarcoplasmic reticulum Ca2+-ATPase activity and cardiac function in rats with diabetes mellitus (DM) mediated by recombinant adeno-associated virus (rAAV) vector. Methods Six weeks after the induction of DM by streptozotocin injected intraperitoneally, the rats were divided into three groups, namely: DM-rAAV-asPLB group, DM-saline group and DM group (control group). The rats in the DM-rAAV-asPLB group were intramyocardially injected with rAAV-asPLB, the rats in the DM-saline group were injected with saline, and those in the control group did not receive any treatment. Six weeks after gene transfer, the expressions of PLB protein and PLB phosphorylation were detected by Western-blot, while the activity of sarcoplasmic reticulum (SR) Ca2+-ATPase and left ventricular function were measured. Results The PLB protein expression level was significantly higher whereas the PLB phosphorylation, SR Ca2+-ATPase activity and left ventricular function were significantly lower in the DM-saline group than in the control group. No significant difference was found in PLB protein expression level, PLB phosphorylation or SR Ca2+-ATPase activity between the DM-rAAV-asPLB group and the control group. The left ventricular function in the DM-rAAV-asPLB group was poorer than in the control group and was better than in the DM-saline group. Conclusion rAAV-asPLB can down-regulate PLB protein expression and up-regulate PLB phosphorylation and SR Ca2+ -ATPase activity, thus contributing to the improvement of in vivo left ventricular function.
基金the National Natural Science Foundation for Distinguished Young Scholars of China(Grant No.82125004)the Shenzhen Science and Technology Innovation Commission(Grant No.JCYJ20220818103414030)+2 种基金the National Natural Science Foundation of China(Grant No.82300397)the key project of Shenzhen Basic Research Program(Natural Science Foundation of Shenzhen,Grant No.20220241)the Program for Guangdong Introducing Innovative and Enterpreneurial Teams(Grant No.2019ZT08Y481).
文摘This study aimed to determine the prevalence and clinical features of Arrhythmogenic Right Ventricular Cardiomyopathy(ARVC)caused by pathogenic mutations in the Phospholamban(PLN)gene.The study included 170 patients who had a confrmed diagnosis of ARVC and underwent PLN genetic screening using next-generation sequencing.The fndings of this study provide valuable insights into the association between PLN mutations and ARVC,which can aid in the development of more efective diagnostic and treatment strategies for ARVC patients.Out of the patients evaluated,six had a rare pathogenic mutation in PLN with the same p.R14del variant.Family screening revealed that heterozygous carriers of p.R14del exhibited a defnite ARVC phenotype.In clinical studies,individuals with the p.R14del mutation experienced a similar rate of malignant arrhythmia events as those with classic desmosome mutations.After adjusting for covariates,individuals with PLN mutations had a two point one seven times greater likelihood of experiencing transplant-related risks compared to those who did not possess PLN mutations(95%CI 1.08–6.82,p=0.035).The accumulation of left ventricular fat and fbers is a pathological marker for ARVC patients with p.R14del mutations.In a cohort of 170 Chinese ARVC patients,three point fve percent of probands had the PLN pathogenic variant(p.R14del)and all were female.Our data shows that PLN-related ARVC patients are at high risk for ventricular arrhythmias and heart failure,which requires clinical diferentiation from classic ARVC.Furthermore,carrying the p.R14del mutation can be an independent prognostic risk factor in ARVC patients.
基金supported by the National Natural Science Foundation of China (20773085 and 30870476)National High-Tech Research and Devel-opment Program of China (2007AA02Z333)Major Chinese National Funding of New Drug Discovery for the Integrated Platform
文摘Phospholamban is an important protein with responsibility for regulating the activity of the sarcoplasmic reticulum Ca2+ pump through reversible phosphorylation.And its three-dimensional structure in living cell has been a focus of attention.In the current case, we summarized the investigations on phospholamban structure, and on this base, employed long time-scale molecular dy-namics simulations to study its structure systematically.The first 22 residues from one chain of phospholamban in bellflower structure determined by NMR experiments, together with its phosphorylation at position 16 and mutation at position 9 were picked up as three different systems.By molecular dynamics simulations of 10 ns in the explicit solution surroundings, it was found that the 3–15 residues of the original structure retained their helix structures, while the phosphorylation and mutation had less probability to form helix structures.These structural changes might result in inhibition decrease to the sarcoplasmic reticulum Ca2+ pump, which is in accordance with previous experimental results.
文摘目的:研究扩心方对阿霉素诱导扩张型心肌病模型大鼠心功能、心肌损伤的保护作用,探讨扩心方作用机制。方法:50只Wistar大鼠腹腔注射阿霉素6周建立DCM模型,开始造模4周后随机分为模型组,扩心方高、中、低剂量组,卡托普利组(每组各10只),连续灌胃给药四周,另设10只正常对照。正常组和模型组以生理盐水灌胃。治疗结束后,对各组大鼠进行心脏超声检查,病理形态学检测,以及用反转录-聚合酶链反应和蛋白质免疫印迹法分别检测心肌组织中SERCA2α、PLB的m RNA和蛋白含量。结果:与正常对照组相比,模型组心功能减弱,出现心肌病理损害,SERCA2α与PLB表达异常;与模型组相比,扩心方组可以改善DCM大鼠的心功能、心肌病理学形态,同时,提高SERCA2αm RNA和蛋白表达、抑制PLB m RNA和蛋白表达。结论:扩心方能提高DCM大鼠的左心室射血分数和缩短率,减轻心肌损伤,改善心功能,同时调控DCM大鼠SERCA2α、PLB的表达。提示扩心方改善DCM大鼠心功能的作用机制,可能与纠正心肌肌浆网钙调控相关蛋白-SERCA2α、PLB的异常表达相关。
