Pub Date : 2023-04-29DOI: 10.1007/s12104-023-10132-8
Bruno Hargittay, Konstantin S. Mineev, Christian Richter, Sridhar Sreeramulu, Hendrik R.A. Jonker, Krishna Saxena, Harald Schwalbe
The splicing isoform b of human fibroblast growth factor 8 (FGF8b) is an important regulator of brain embryonic development. Here, we report the almost complete NMR chemical shift assignment of the backbone and aliphatic side chains of FGF8b. Obtained chemical shifts are in good agreement with the previously reported X-ray data, excluding the N-terminal gN helix, which apparently forms only in complex with the receptor. The reported data provide an NMR starting point for the investigation of FGF8b interaction with its receptors and with potential drugs or inhibitors.
{"title":"NMR resonance assignment of a fibroblast growth factor 8 splicing isoform b","authors":"Bruno Hargittay, Konstantin S. Mineev, Christian Richter, Sridhar Sreeramulu, Hendrik R.A. Jonker, Krishna Saxena, Harald Schwalbe","doi":"10.1007/s12104-023-10132-8","DOIUrl":"10.1007/s12104-023-10132-8","url":null,"abstract":"<div><p>The splicing isoform b of human fibroblast growth factor 8 (FGF8b) is an important regulator of brain embryonic development. Here, we report the almost complete NMR chemical shift assignment of the backbone and aliphatic side chains of FGF8b. Obtained chemical shifts are in good agreement with the previously reported X-ray data, excluding the N-terminal gN helix, which apparently forms only in complex with the receptor. The reported data provide an NMR starting point for the investigation of FGF8b interaction with its receptors and with potential drugs or inhibitors.</p></div>","PeriodicalId":492,"journal":{"name":"Biomolecular NMR Assignments","volume":null,"pages":null},"PeriodicalIF":0.9,"publicationDate":"2023-04-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s12104-023-10132-8.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"5589130","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-04-26DOI: 10.1007/s12104-023-10124-8
Agnes Adler, Lenette F. Kjaer, J. Wouter Beugelink, Marc Baldus, Hugo van Ingen
The microtubule-associated protein 7 (MAP7) is a protein involved in cargo transport along microtubules (MTs) by interacting with kinesin-1 through the C-terminal kinesin-binding domain. Moreover, the protein is reported to stabilize MT, thereby playing a key role in axonal branch development. An important element for this latter function is the 112 amino-acid long N-terminal microtubule-binding domain (MTBD) of MAP7. Here we report NMR backbone and side-chain assignments that suggest a primarily alpha-helical secondary fold of this MTBD in solution. The MTBD contains a central long α-helical segment that includes a short four-residue ‘hinge’ sequence with decreased helicity and increased flexibility. Our data represent a first step towards analysing the complex interaction of MAP7 with MTs at an atomic level via NMR spectroscopy.
{"title":"Resonance assignments of the microtubule-binding domain of the microtubule-associated protein 7 (MAP7)","authors":"Agnes Adler, Lenette F. Kjaer, J. Wouter Beugelink, Marc Baldus, Hugo van Ingen","doi":"10.1007/s12104-023-10124-8","DOIUrl":"10.1007/s12104-023-10124-8","url":null,"abstract":"<div><p>The microtubule-associated protein 7 (MAP7) is a protein involved in cargo transport along microtubules (MTs) by interacting with kinesin-1 through the C-terminal kinesin-binding domain. Moreover, the protein is reported to stabilize MT, thereby playing a key role in axonal branch development. An important element for this latter function is the 112 amino-acid long N-terminal microtubule-binding domain (MTBD) of MAP7. Here we report NMR backbone and side-chain assignments that suggest a primarily alpha-helical secondary fold of this MTBD in solution. The MTBD contains a central long α-helical segment that includes a short four-residue ‘hinge’ sequence with decreased helicity and increased flexibility. Our data represent a first step towards analysing the complex interaction of MAP7 with MTs at an atomic level via NMR spectroscopy.</p></div>","PeriodicalId":492,"journal":{"name":"Biomolecular NMR Assignments","volume":null,"pages":null},"PeriodicalIF":0.9,"publicationDate":"2023-04-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s12104-023-10124-8.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"4997779","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-04-07DOI: 10.1007/s12104-023-10125-7
Aritra Bej, James B. Ames
N-methyl-D-aspartate receptors (NMDARs) consist of glycine-binding GluN1 and glutamate-binding GluN2 subunits that form tetrameric ion channels. NMDARs in the neuronal post-synaptic membrane are important for controlling neuroplasticity and synaptic transmission in the brain. Calmodulin (CaM) binds to the cytosolic C0 domains of both GluN1 (residues 841–865) and GluN2 (residues 1004–1024) that may play a role in the Ca2+-dependent desensitization of NMDAR channels. Mutations that disrupt Ca2+-dependent desensitization of NMDARs are linked to Alzheimer’s disease, depression, stroke, epilepsy, and schizophrenia. NMR chemical shift assignments are reported here for Ca2+-saturated CaM bound to the GluN2A C0 domain of NMDAR (BMRB no. 51821).
n -甲基- d -天冬氨酸受体(NMDARs)由甘氨酸结合GluN1和谷氨酸结合GluN2亚基组成,形成四聚体离子通道。神经元突触后膜中的NMDARs对控制大脑神经可塑性和突触传递具有重要意义。钙调蛋白(CaM)结合GluN1(残基841-865)和GluN2(残基1004-1024)的细胞质C0结构域,可能在NMDAR通道的Ca2+依赖性脱敏中发挥作用。破坏NMDARs Ca2+依赖性脱敏的突变与阿尔茨海默病、抑郁症、中风、癫痫和精神分裂症有关。本文报道了Ca2+饱和CaM与NMDAR (BMRB no. 1)的GluN2A C0结构域结合的NMR化学位移分配。51821)。
{"title":"Chemical shift assignments of calmodulin bound to a cytosolic domain of GluN2A (residues 1004–1024) from the NMDA receptor","authors":"Aritra Bej, James B. Ames","doi":"10.1007/s12104-023-10125-7","DOIUrl":"10.1007/s12104-023-10125-7","url":null,"abstract":"<div><p>N-methyl-D-aspartate receptors (NMDARs) consist of glycine-binding GluN1 and glutamate-binding GluN2 subunits that form tetrameric ion channels. NMDARs in the neuronal post-synaptic membrane are important for controlling neuroplasticity and synaptic transmission in the brain. Calmodulin (CaM) binds to the cytosolic C0 domains of both GluN1 (residues 841–865) and GluN2 (residues 1004–1024) that may play a role in the Ca<sup>2+</sup>-dependent desensitization of NMDAR channels. Mutations that disrupt Ca<sup>2+</sup>-dependent desensitization of NMDARs are linked to Alzheimer’s disease, depression, stroke, epilepsy, and schizophrenia. NMR chemical shift assignments are reported here for Ca<sup>2+</sup>-saturated CaM bound to the GluN2A C0 domain of NMDAR (BMRB no. 51821).</p></div>","PeriodicalId":492,"journal":{"name":"Biomolecular NMR Assignments","volume":null,"pages":null},"PeriodicalIF":0.9,"publicationDate":"2023-04-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s12104-023-10125-7.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"4606241","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-04-06DOI: 10.1007/s12104-023-10127-5
Gwen R. Buel, Xiang Chen, Olumide Kayode, Anthony Cruz, Kylie J. Walters
UBQLN1 functions in autophagy and proteasome-mediated protein degradation. It contains an N-terminal ubiquitin-like domain (UBL), a C-terminal ubiquitin-associated domain (UBA), and a flexible central region which functions as a chaperone to prevent protein aggregation. Here, we report the 1H, 15N, and 13C resonance assignments for the backbone (NH, N, C’, Cα, and Hα) and sidechain Cβ atoms of the UBQLN1 UBA and an N-terminally adjacent segment called the UBA-adjacent domain (UBAA). We find a subset of the resonances corresponding to the UBAA to have concentration-dependent chemical shifts, likely due to self-association. We also find the backbone amide nitrogen of T572 to be shifted upfield relative to the average value for a threonine amide nitrogen, a phenomenon likely caused by T572 Hγ1 engagement in a hydrogen bond with adjacent backbone carbonyl atoms. The assignments described in this manuscript can be used to study the protein dynamics of the UBQLN1 UBA and UBAA as well as the interaction of these domains with other proteins.
UBQLN1在自噬和蛋白酶体介导的蛋白质降解中起作用。它包含一个n端泛素样结构域(UBL),一个c端泛素相关结构域(UBA)和一个灵活的中心区域,作为伴侣防止蛋白质聚集。在这里,我们报道了UBQLN1 UBA的主链(NH, N, C ', Cα和Hα)和侧链Cβ原子以及称为UBA相邻结构域(UBAA)的N端相邻片段的1H, 15N和13C共振分配。我们发现与UBAA对应的共振子集具有浓度依赖的化学位移,可能是由于自关联。我们还发现T572的主酰胺氮相对于苏氨酸酰胺氮的平均值向上移动,这一现象可能是由于T572的h - γ1与相邻的主羰基原子的氢键作用引起的。本文所描述的结构域可用于研究UBQLN1 UBA和UBAA的蛋白质动力学以及这些结构域与其他蛋白质的相互作用。
{"title":"1H, 15N, 13C backbone and Cβ resonance assignments for UBQLN1 UBA and UBAA domains","authors":"Gwen R. Buel, Xiang Chen, Olumide Kayode, Anthony Cruz, Kylie J. Walters","doi":"10.1007/s12104-023-10127-5","DOIUrl":"10.1007/s12104-023-10127-5","url":null,"abstract":"<div><p>UBQLN1 functions in autophagy and proteasome-mediated protein degradation. It contains an N-terminal ubiquitin-like domain (UBL), a C-terminal ubiquitin-associated domain (UBA), and a flexible central region which functions as a chaperone to prevent protein aggregation. Here, we report the <sup>1</sup>H, <sup>15</sup>N, and <sup>13</sup>C resonance assignments for the backbone (<sup>N</sup>H, N, C’, Cα, and Hα) and sidechain Cβ atoms of the UBQLN1 UBA and an N-terminally adjacent segment called the UBA-adjacent domain (UBAA). We find a subset of the resonances corresponding to the UBAA to have concentration-dependent chemical shifts, likely due to self-association. We also find the backbone amide nitrogen of T572 to be shifted upfield relative to the average value for a threonine amide nitrogen, a phenomenon likely caused by T572 Hγ1 engagement in a hydrogen bond with adjacent backbone carbonyl atoms. The assignments described in this manuscript can be used to study the protein dynamics of the UBQLN1 UBA and UBAA as well as the interaction of these domains with other proteins.</p></div>","PeriodicalId":492,"journal":{"name":"Biomolecular NMR Assignments","volume":null,"pages":null},"PeriodicalIF":0.9,"publicationDate":"2023-04-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s12104-023-10127-5.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"4238043","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-04-06DOI: 10.1007/s12104-023-10126-6
Rahul Yadav, Tanveer Shaikh, Suhas Tikole, Andrew B. Herr, Nicholas C. Fitzkee
Staphylococcus epidermidis is the leading causative agent for hospital-acquired infections, especially device-related infections, due to its ability to form biofilms. The accumulation-associated protein (Aap) of S. epidermidis is primarily responsible for biofilm formation and consists of two domains, A and B. It was found that the A domain is responsible for the attachment to the abiotic/biotic surface, whereas the B domain is responsible for the accumulation of bacteria during biofilm formation. One of the parts of the A domain is the Aap lectin, which is a carbohydrate-binding domain having 222 amino acids in its structure. Here we report the near complete backbone chemical shift assignments for the lectin domain, as well as its predicted secondary structure. This data will provide a platform for future NMR studies to explore the role of lectin in biofilm formation.
{"title":"1H, 15N, and 13C chemical shift backbone resonance NMR assignment of the accumulation-associated protein (Aap) lectin domain from Staphylococcus epidermidis","authors":"Rahul Yadav, Tanveer Shaikh, Suhas Tikole, Andrew B. Herr, Nicholas C. Fitzkee","doi":"10.1007/s12104-023-10126-6","DOIUrl":"10.1007/s12104-023-10126-6","url":null,"abstract":"<div><p><i>Staphylococcus epidermidis</i> is the leading causative agent for hospital-acquired infections, especially device-related infections, due to its ability to form biofilms. The accumulation-associated protein (Aap) of <i>S. epidermidis</i> is primarily responsible for biofilm formation and consists of two domains, A and B. It was found that the A domain is responsible for the attachment to the abiotic/biotic surface, whereas the B domain is responsible for the accumulation of bacteria during biofilm formation. One of the parts of the A domain is the Aap lectin, which is a carbohydrate-binding domain having 222 amino acids in its structure. Here we report the near complete backbone chemical shift assignments for the lectin domain, as well as its predicted secondary structure. This data will provide a platform for future NMR studies to explore the role of lectin in biofilm formation.</p></div>","PeriodicalId":492,"journal":{"name":"Biomolecular NMR Assignments","volume":null,"pages":null},"PeriodicalIF":0.9,"publicationDate":"2023-04-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s12104-023-10126-6.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"4238070","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-03-01DOI: 10.1007/s12104-023-10123-9
Tsega L. Solomon, Kinlin Chao, Genevieve Gingras, Yves Aubin, William B. O’Dell, John P. Marino, Robert G. Brinson
The monoclonal antibody (mAb) protein class has become a primary therapeutic platform for the production of new life saving drug products. MAbs are comprised of two domains: the antigen-binding fragment (Fab) and crystallizable fragment (Fc). Despite the success in the clinic, NMR assignments of the complete Fab domain have been elusive, in part due to problems in production of properly folded, triply-labeled 2H,13C,15N Fab domain. Here, we report the successful recombinant expression of a triply-labeled Fab domain, derived from the standard IgG1κ known as NISTmAb, in yeast. Using the 2H,13C,15N Fab domain, we assigned 94% of the 1H, 13C, and 15N backbone atoms.
{"title":"Backbone NMR assignment of the yeast expressed Fab fragment of the NISTmAb reference antibody","authors":"Tsega L. Solomon, Kinlin Chao, Genevieve Gingras, Yves Aubin, William B. O’Dell, John P. Marino, Robert G. Brinson","doi":"10.1007/s12104-023-10123-9","DOIUrl":"10.1007/s12104-023-10123-9","url":null,"abstract":"<div><p>The monoclonal antibody (mAb) protein class has become a primary therapeutic platform for the production of new life saving drug products. MAbs are comprised of two domains: the antigen-binding fragment (Fab) and crystallizable fragment (Fc). Despite the success in the clinic, NMR assignments of the complete Fab domain have been elusive, in part due to problems in production of properly folded, triply-labeled <sup>2</sup>H,<sup>13</sup>C,<sup>15</sup>N Fab domain. Here, we report the successful recombinant expression of a triply-labeled Fab domain, derived from the standard IgG1κ known as NISTmAb, in yeast. Using the <sup>2</sup>H,<sup>13</sup>C,<sup>15</sup>N Fab domain, we assigned 94% of the <sup>1</sup>H, <sup>13</sup>C, and <sup>15</sup>N backbone atoms.</p></div>","PeriodicalId":492,"journal":{"name":"Biomolecular NMR Assignments","volume":null,"pages":null},"PeriodicalIF":0.9,"publicationDate":"2023-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s12104-023-10123-9.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"4042515","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-02-10DOI: 10.1007/s12104-023-10120-y
Julian C.-H. Chen, Marco Tonelli, Penelope Anderson, Ryszard Michalczyk, Marc-Michael Blum, Robert F. Williams
NMR chemical shift assignments are reported for backbone (15N, 1H) and partial side chain (13Cα and β, side chain 1H) atoms of diisopropyl fluorophosphatase (DFPase), a calcium-dependent phosphotriesterase capable of hydrolyzing phosphorus – fluorine bonds in a variety of toxic organophosphorus compounds. Analysis of residues lining the active site of DFPase highlight a number of residues whose chemical shifts can be used as a diagnostic of binding and detection of organophosphorus compounds.
{"title":"Backbone and side chain chemical shift assignment of diisopropyl fluorophosphatase (DFPase) from Loligo vulgaris, an organophosphorus-degrading enzyme","authors":"Julian C.-H. Chen, Marco Tonelli, Penelope Anderson, Ryszard Michalczyk, Marc-Michael Blum, Robert F. Williams","doi":"10.1007/s12104-023-10120-y","DOIUrl":"10.1007/s12104-023-10120-y","url":null,"abstract":"<div><p>NMR chemical shift assignments are reported for backbone (<sup>15</sup>N, <sup>1</sup>H) and partial side chain (<sup>13</sup>Cα and β, side chain <sup>1</sup>H) atoms of diisopropyl fluorophosphatase (DFPase), a calcium-dependent phosphotriesterase capable of hydrolyzing phosphorus – fluorine bonds in a variety of toxic organophosphorus compounds. Analysis of residues lining the active site of DFPase highlight a number of residues whose chemical shifts can be used as a diagnostic of binding and detection of organophosphorus compounds.</p></div>","PeriodicalId":492,"journal":{"name":"Biomolecular NMR Assignments","volume":null,"pages":null},"PeriodicalIF":0.9,"publicationDate":"2023-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://link.springer.com/content/pdf/10.1007/s12104-023-10120-y.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"4417636","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-02-09DOI: 10.1007/s12104-023-10122-w
Anamika Sulekha, Michael J. Osborne, Jadwiga Gasiorek, Katherine L. B. Borden
UDP-glucuronosyltransferases are the principal enzymes involved in the glucuronidation of metabolites and xenobiotics for physiological clearance in humans. Though glucuronidation is an indispensable process in the phase II metabolic pathway, UGT-mediated glucuronidation of most prescribed drugs (> 55%) and clinical evidence of UGT-associated drug resistance are major concerns for therapeutic development. While UGTs are highly conserved enzymes, they manifest unique substrate and inhibitor specificity which is poorly understood given the dearth of experimentally determined full-length structures. Such information is important not only to conceptualize their specificity but is central to the design of inhibitors specific to a given UGT in order to avoid toxicity associated with pan-UGT inhibitors. Here, we provide the 1H, 13C and 15N backbone (~ 90%) and sidechain (~ 62%) assignments for the C-terminal domain of UGT2B17, which can be used to determine the molecular binding sites of inhibitor and substrate, and to understand the atomic basis for inhibitor selectivity between UGT2B17 and other members of the UGT2B subfamily. Given the physiological relevance of UGT2B17 in the elimination of hormone-based cancer drugs, these assignments will contribute towards dissecting the structural basis for substrate specificity, selective inhibitor recognition and other aspects of enzyme activity with the goal of selectively overcoming glucuronidation-based drug resistance.
{"title":"1H, 13C, 15N Backbone and sidechain chemical shift assignments of the C-terminal domain of human UDP-glucuronosyltransferase 2B17 (UGT2B17-C)","authors":"Anamika Sulekha, Michael J. Osborne, Jadwiga Gasiorek, Katherine L. B. Borden","doi":"10.1007/s12104-023-10122-w","DOIUrl":"10.1007/s12104-023-10122-w","url":null,"abstract":"<div><p>UDP-glucuronosyltransferases are the principal enzymes involved in the glucuronidation of metabolites and xenobiotics for physiological clearance in humans. Though glucuronidation is an indispensable process in the phase II metabolic pathway, UGT-mediated glucuronidation of most prescribed drugs (> 55%) and clinical evidence of UGT-associated drug resistance are major concerns for therapeutic development. While UGTs are highly conserved enzymes, they manifest unique substrate and inhibitor specificity which is poorly understood given the dearth of experimentally determined full-length structures. Such information is important not only to conceptualize their specificity but is central to the design of inhibitors specific to a given UGT in order to avoid toxicity associated with pan-UGT inhibitors. Here, we provide the <sup>1</sup>H, <sup>13</sup>C and <sup>15</sup>N backbone (~ 90%) and sidechain (~ 62%) assignments for the C-terminal domain of UGT2B17, which can be used to determine the molecular binding sites of inhibitor and substrate, and to understand the atomic basis for inhibitor selectivity between UGT2B17 and other members of the UGT2B subfamily. Given the physiological relevance of UGT2B17 in the elimination of hormone-based cancer drugs, these assignments will contribute towards dissecting the structural basis for substrate specificity, selective inhibitor recognition and other aspects of enzyme activity with the goal of selectively overcoming glucuronidation-based drug resistance.</p></div>","PeriodicalId":492,"journal":{"name":"Biomolecular NMR Assignments","volume":null,"pages":null},"PeriodicalIF":0.9,"publicationDate":"2023-02-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"4382835","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The Endosomal Sorting Complex Required for Transport (ESCRT) pathway, through inverse topology membrane remodeling, is involved in many biological functions, such as ubiquitinated membrane receptor trafficking and degradation, multivesicular bodies (MVB) formation and cytokinesis. Dysfunctions in ESCRT pathway have been associated to several human pathologies, such as cancers and neurodegenerative diseases. The ESCRT machinery is also hijacked by many enveloped viruses to bud away from the plasma membrane of infected cells. Human tumor susceptibility gene 101 (Tsg101) protein is an important ESCRT-I complex component. The structure of the N-terminal ubiquitin E2 variant (UEV) domain of Tsg101 (Tsg101-UEV) comprises an ubiquitin binding pocket next to a late domain [P(S/T)AP] binding groove. These two binding sites have been shown to be involved both in the physiological roles of ESCRT-I and in the release of the viral particles, and thus are attractive targets for antivirals. The structure of the Tsg101-UEV domain has been characterized, using X-ray crystallography or NMR spectroscopy, either in its apo-state or bound to ubiquitin or late domains. In this study, we report the backbone NMR resonance assignments, including the proline signals, of the apo human Tsg101-UEV domain, that so far was not publicly available. These data, that are in good agreement with the crystallographic structure of Tsg101-UEV domain, can therefore be used for further NMR studies, including protein-protein interaction studies and drug discovery.
ESCRT (Endosomal Sorting Complex Required for Transport)通路通过逆拓扑膜重构参与多种生物学功能,如泛素化膜受体的转运和降解、多泡体(multivesular bodies, MVB)的形成和细胞分裂。ESCRT通路功能障碍与多种人类病理相关,如癌症和神经退行性疾病。ESCRT机制也被许多包膜病毒劫持,从被感染细胞的质膜上萌芽。人肿瘤易感基因101 (Tsg101)蛋白是ESCRT-I复合体的重要组成部分。Tsg101的n端泛素E2变体(UEV)结构域(Tsg101-UEV)的结构包括一个泛素结合口袋,旁边是一个晚期结构域[P(S/T)AP]结合槽。这两个结合位点已被证明参与ESCRT-I的生理作用和病毒颗粒的释放,因此是抗病毒药物的有吸引力的靶点。利用x射线晶体学或核磁共振光谱对Tsg101-UEV结构域的载脂蛋白态或与泛素或晚期结构域结合的结构进行了表征。在这项研究中,我们报道了迄今为止尚未公开的载子人类Tsg101-UEV结构域的主干核磁共振分配,包括脯氨酸信号。这些数据与Tsg101-UEV结构域的晶体结构非常吻合,因此可以用于进一步的核磁共振研究,包括蛋白质-蛋白质相互作用研究和药物发现。
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Pub Date : 2023-02-05DOI: 10.1007/s12104-023-10121-x
Aritra Bej, James B. Ames
Neuroplasticity and synaptic transmission in the brain are regulated by N-methyl-D-aspartate receptors (NMDARs) that consist of hetero-tetrameric combinations of the glycine-binding GluN1 and glutamate-binding GluN2 subunits. Calmodulin (CaM) binds to the cytosolic C0 domain of GluN1 (residues 841–865) that may play a role in the Ca2+-dependent inactivation (CDI) of NMDAR channel activity. Dysregulation of NMDARs are linked to various neurological disorders, including Alzheimer’s disease, depression, stroke, epilepsy, and schizophrenia. Here, we report complete NMR chemical shift assignments of Ca2+-saturated CaM bound to the GluN1 C0 domain of the human NMDAR (BMRB no. 51715).
大脑中的神经可塑性和突触传递由n -甲基- d -天冬氨酸受体(NMDARs)调节,NMDARs由甘氨酸结合GluN1和谷氨酸结合GluN2亚基的异四聚体组合组成。钙调蛋白(CaM)与GluN1的胞质C0结构域(残基841-865)结合,可能在NMDAR通道活性的Ca2+依赖性失活(CDI)中发挥作用。NMDARs的失调与各种神经系统疾病有关,包括阿尔茨海默病、抑郁症、中风、癫痫和精神分裂症。在这里,我们报告了Ca2+饱和CaM结合到人类NMDAR (BMRB no. 1)的GluN1 C0结构域的完整NMR化学位移分配。51715)。
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