共查询到20条相似文献,搜索用时 31 毫秒
1.
2.
J. Graham Martin 《BMJ (Clinical research ed.)》1935,2(3906):970-971
3.
4.
《Journal of molecular biology》2019,431(14):2554-2566
Proton transfer through membrane-bound ion channels is mediated by both water and polar residues of proteins, but the detailed molecular mechanism is challenging to determine. The tetrameric influenza A and B virus M2 proteins form canonical proton channels that use an HxxxW motif for proton selectivity and gating. The BM2 channel also contains a second histidine (His), H27, equidistant from the gating tryptophan, which leads to a symmetric H19xxxW23xxxH27 motif. The proton-dissociation constants (pKa's) of H19 in BM2 were found to be much lower than the pKa's of H37 in AM2. To determine if the lower pKa's result from H27-facilitated proton dissociation of H19, we have now investigated a H27A mutant of BM2 using solid-state NMR. 15N NMR spectra indicate that removal of the second histidine converted the protonation and tautomeric equilibria of H19 to be similar to the H37 behavior in AM2, indicating that the peripheral H27 is indeed the origin of the low pKa's of H19 in wild-type BM2. Measured interhelical distances between W23 sidechains indicate that the pore constriction at W23 increases with the H19 tetrad charge but is independent of the H27A mutation. These results indicate that H27 both accelerates proton dissociation from H19 to increase the inward proton conductance and causes the small reverse conductance of BM2. The proton relay between H19 and H27 is likely mediated by the intervening gating tryptophan through cation–π interactions. This relayed proton transfer may exist in other ion channels and has implications for the design of imidazole-based synthetic proton channels. 相似文献
5.
6.
目的:对His/GST-HDAC1在大肠杆菌BL-21中的表达进行研究。方法:HDAC1的完整基因片断被克隆到pColdⅠ载体和pGEX载体上,并在其N末端分别联有His和GST;采用大肠杆菌BL21对HDAC1进行表达;采用亲和色谱对HDAC1进行纯化;用SDS-PAGE和蛋白质印迹来验证表达和纯化效果;对HDAC1活性进行测定。结果:多数HDAC1存在于大肠杆菌BL-21细胞裂解液的沉淀组分和纯化过程中的未结合组分中,小部分HDAC1可从细胞裂解液的上清液中得以纯化,但未显现出酶活性。用FPLC对HDAC1进行进一步分离,结果表明,HDAC1发生了分子聚集,使得它们的分子量大于正常分子量。结论:活性His/GST-HDAC1不能用大肠杆菌BL21成功表达。 相似文献
7.
NMT在大肠杆菌中的His6融合表达及其纯化研究 总被引:2,自引:0,他引:2
将啤酒酵母NMT基因以N端融合6个组氨酸的形式在E.coliBL21(DE3)中进行了IPTG诱导的表达研究。SDS-PAGE分析确定有与His。-rNMT理论分子量一致的诱导表达条带,其表达量占全菌蛋白的10%左右;表达产物性质分析表明His-rNMT主要以可溶形式存在。在此基础上不受利用固定化金属离子配体亲和层析一步纯化目的蛋白,纯度可达95%以上。体外标脾性是His6-rNMT具有与成熟NM 相似文献
8.
Summary A new chain mutant Hb-Sinai 47
His is described. The aminoacid composition of all tryptic peptides has been determined, with the exception of the insoluble core. In the fingerprint peptide T 6 which normally migrates between T 3 and T 1, moves now between T 3 and T 7. The aminoacid composition of peptide T 6 indicates a change in the aminoacid composition from Asp- to His+ in position 47.This work was supported by Grant No. GM 13714 U.S.P.H.S. 相似文献
9.
Duilio F. Raffa Rodolfo Gómez-Balderas Patrick Brunelle Gail A. Rickard Arvi Rauk 《Journal of biological inorganic chemistry》2005,10(8):887-902
Two of the defining hallmarks of Alzheimer’s disease (AD) are deposits of the β-amyloid peptide, Aβ, and the generation of
reactive oxygen species, both of which may be due to the Aβ peptide coordinating metal ions. The Cu2+ concentrations in cores of senile plaques are significantly elevated in AD patients. Experimental results indicate that Aβ1–42
in particular has a very high affinity for Cu2+, and that His13 and His14 are the two most firmly established ligands in the coordination sphere of the copper ion. Quantum
chemical calculations using the unrestricted B3LYP hybrid density functional method with the 6–31G(d) basis set were performed
for geometries, zero point energies and thermochemistry. The effects of solvation were accommodated using the CPCM method.
The enthalpies were calculated with the 6–311+G(2df,2p) basis set. Calculations show that when Cu(H2O)42+ combines with the model compound 1 (3-(1H-imidazol-5-yl)-N-[2-(1H-imidazol-5-yl)ethyl] propanamide) in the aqueous phase, the most stable binding site involves the Nπ atoms of His13 and His14
as well as the carbonyl of the intervening backbone amide group. These structures are fairly rigid and the implications for
conformational changes to the Aβ backbone are discussed. In solution at pH=7, Cu2+ promotes the deprotonation and involvement in the binding of the backbone amide nitrogen in a β-sheet like structure. This
geometry does not induce strain in the peptide backbone, making it the most likely representation of that portion of the Cu2+–Aβ complex monomer in aqueous solution. 相似文献
10.
S. Salvadori R. Guerrini V. Forlani S. D. Bryant M. Attila L. H. Lazarus 《Amino acids》1994,7(3):291-304
Summary Analysis of deltorphin A position 4 analogues included: backbone constrained N
MeHis, spinacine (Spi), N
MePhe and the tetrahydroisoquinoline-3-carboxylic acid (Tic); spatially confined side-chain (Phg); and imidazole alkylation ofl- andd-His4 enantiomers. High selectivity was lost with the following replacements: N
MeHis4, N
MePhe4 and Phg4 reduced binding and the constrained residues also increasedµ binding; ring closure between the side-chain and amino group to yield Spi4 or Tic4 increasedµ affinity. Imidazole methylation of His4 marginally affected opioid binding and doubled selectivity; alkylatedd-His4-derivatives generally maintained selectivity in spite of decreased affinities. Thus, His4 imidazole preserves selectivity by facilitating high binding and by repulsion at theµ receptor. Several low energy conformers of deltorphin A indicated that the His4 imidazole preferred a spatial orientation parallel to the phenolic side-chain of Tyr1 suggestive that this conformation might contribute to high affinity and selectivity. 相似文献
11.
12.
Rita Meurisse Robert Brasseur Annick Thomas 《Biochimica et Biophysica Acta - Proteins and Proteomics》2003,1649(1):85-96
Several studies have analysed aromatic interactions, involving mostly phenylalanine, tyrosine and tryptophan. Only a few studies have considered histidine as an interacting aromatic residue. An extensive analysis of aromatic His–X interactions is performed here on a data set of 593 PDB structures: 68% of the histidine are involved in aromatic pairs and 1271 non-redundant His–X pairs were analysed. Thirty percent of these pairs involve an aromatic partner less than 6 apart in the sequence. These near-sequence pairs correspond to conformations which stabilise secondary structures, mainly α-helices when the residues are 4 apart and β-strands when they are 2 apart in the sequence. The partners of the other His–X pairs (887, 70%) are more than 5 apart in the sequence. Of these far-sequence pairs, 35% bridge beta strands and only 9% helices. The near-sequence pairs are sterically constrained as supported by conformer distribution. The X partners of far-sequence His–X pairs are mainly “above” the histidine ring with tilted and normal rings, corresponding to a “T shape; face to edge” orientation. Phenylalanine, the only aromatic residue with no heteroatom, is a disfavoured partner, whereas histidine is the preferred one. Heteroatom–heteroatom interactions are favoured in near-sequence as well as in far-sequence His–His, His–Trp and His–Tyr pairs. 相似文献
13.
大肠杆菌His6融合表达载体及其表达产物的一步变化 总被引:10,自引:0,他引:10
构建了T7启动子控制下的融合表达载体,使外源蛋白以N端与6个连续的组氨酸残基融合的形式表达。这些载体含有强T7启动子,终止子,翻译起始区,多克隆位点以及噬菌体f1复制起始区,使外源基因不仅可以得到高效表达而且可以利用helper phage包装单链DNA,从而不需亚克隆玉可以直接进行测序及点突变。 相似文献
14.
15.
SDS—PAPGE法测定His—tag融合蛋白分子量产生偏差的原因 总被引:5,自引:0,他引:5
His-tag/Ni-NTA系统是新发展起来的一个亲和纯化重组蛋白有用工具,现常用于基因编码产物的特性研究中。SDS-PAGE是实验室测定蛋白质分子量通常采用的方法,而许多实验室用此方法检测His-tag融合蛋白时却常发现测得的分子量偏大,产生偏差的原因尚未阐明。为弄清这一问题,本实验室在研究一个His-tag融合蛋白P73-His时,首先用SDS-PAGE法测得其分子量确实比理论计算值大,然后对 相似文献
16.
David L. Hull 《Biology & philosophy》2001,16(3):423-435
In this paper I trace Michael Ruse's Booknotes from the first volumeof Biology and Philosophy in 1986 to the present. I deal withboth the style and the content of these booknotes. Ruse paid specialattention to authors outside of the traditional English axis as wellas to feminist writers. He complained that too much attention wasbeing paid to certain topics (e.g., evolutionary ethics, evolutionaryepistemology, the species problem and reduction) while other, moreimportant topics were all but ignored (e.g., natural selection,population genetics, levels of selection and extraterrestrial life).He also dealt with the Darwin Industry. Creationism, his love-haterelationships with several authors and his undiluted love of CharlesDickens. 相似文献
17.
In 1849, Isaac Lea named Sauropus primaevus for footprints from Mount Carbon, Pennsylvania, USA, then the oldest fossil vertebrate footprints reported. In 1902, O. P. Hay constructed a new ichnogenus Palaeosauropus for this ichnospecies. Palaeosauropus has been one of the most frequently reported Mississippian footprint ichnogenera in North America and remains a valid ichnotaxon. The holotype of Palaeosauropus (= “Sauropus”) primaevus (referred to as P. primaevus), consisting of a single manus/pes pair, is described and illustrated in Lea (1853) and is housed at the Academy of Natural Sciences in Philadelphia, Pennsylvania (ANS9752). Lea's large specimen of P. primaevus (approximately 86 cm by 53 cm), that included a trackway of six manus/pes pairs, described and illustrated in 1853 and 1855, was a combination of ANS9752 and a second specimen represented by a plaster cast housed at the National Museum of Natural History (USNM487148). Historical documents and examination of the Mauch Chunk Formation at Mount Carbon, Pennsylvania, enabled the identification of Lea's tracksite, originally reported to be a few hundred feet (about 75 m) from the former Mount Carbon Hotel. Our forensic evidence indicates the type locality for P. primaevus is approximately 90 m south from the southwest corner of Centre and Main Streets in Mount Carbon, Pennsylvania, with geographical coordinates of N 40° 40' 25.7”, W 76° 11' 14.9”. The type locality is within the middle member of the Mauch Chunk Formation, a fluvial sequence of late Mississippian (Visean) Age. 相似文献
18.
Background
How do we bond to one another? While in some species, like humans, physical contact plays a role in the process of attachment, it has been suggested that tactile contact''s value may greatly differ according to the species considered. Nevertheless, grooming is often considered as a pleasurable experience for domestic animals, even though scientific data is lacking. On another hand, food seems to be involved in the creation of most relationships in a variety of species.Methodology/Principal Findings
In this study, we used the horse training context to test the effects of food versus grooming during repeated human-horse interactions. The results reveal that food certainly holds a key role in the attachment process, while tactile contact was here clearly insufficient for bonding to occur.Conclusion/Significance
This study raises important questions on the way tactile contact is perceived, and shows that large inter-species differences are to be expected. 相似文献19.
20.
Duilio F. Raffa Gail A. Rickard Arvi Rauk 《Journal of biological inorganic chemistry》2007,12(2):147-164
A contributing factor to the pathology of Alzheimer's disease is the generation of reactive oxygen species, most probably a consequence of the beta-amyloid (Abeta) peptide coordinating copper ions. Experimental and theoretical results indicate that His13 and His14 are the two most firmly established ligands in the coordination sphere of Cu(II) bound to Abeta. Abeta1-42 is known to reduce Cu(II) to Cu(I). The Abeta-Cu(II) complex has been shown to catalytically generate H(2)O(2) from reducing agents and O(2). Cu(II) in the presence of Abeta has been reported to have a formal reduction potential of +0.72-0.77 V (vs. the standard hydrogen electrode). Quantum chemical calculations using the B3LYP hybrid density functional method with the 6-31G(d) basis set were performed to model the reduction of previously studied Cu(II) complexes representing the His13-His14 portion of Abeta (Raffa et al. in J. Biol. Inorg. Chem. 10:887-902, 2005). The effects of solvation were accommodated using the CPCM method. The most stable complex between Cu(I) and the model compound, 3-(5-imidazolyl)propionylhistamine (1) involves tricoordinated Cu(I) in a distorted-T geometry, with the Npi of both imidazoles as well as the oxygen of the backbone carbonyl bound to copper. This model would be the most likely representation of a Cu(I) binding site for a His-His peptide in aqueous solution. A variety of possible redox processes are discussed. 相似文献