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An attempt has been made to obtain intact individual keratin filaments of various levels from micron cortical, micron macrofibril to nano intermediate filament and polypeptide alpha-helix from the human hair shaft. The feasibility of this initiative has been largely demonstrated by finding that there is a longitudinal seam/zipper on the cuticle of the human hair shaft, which can be unzipped by certain solvents such as performic acid and urea, allowing one to use an anatomical approach to separate intact individual micron/nano filaments. Micron cortical and macrofibril filaments have been obtained. It is also found that the cortical filaments are twisted together to form a yarn, giving rise to the strength for the hair shaft; and that individual cortical filaments are often 2-2 paired in a similar structure to the double helix. 相似文献
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Protein‐engineering of chitosanase from Bacillus sp. MN to alter its substrate specificity 下载免费PDF全文
Eva K. Regel Tobias Weikert Anna Niehues Bruno M. Moerschbacher Ratna Singh 《Biotechnology and bioengineering》2018,115(4):863-873
Partially acetylated chitosan oligosaccharides (paCOS) have various potential applications in agriculture, biomedicine, and pharmaceutics due to their suitable bioactivities. One method to produce paCOS is partial chemical hydrolysis of chitosan polymers, but that leads to poorly defined mixtures of oligosaccharides. However, the effective production of defined paCOS is crucial for fundamental research and for developing applications. A more promising approach is enzymatic depolymerization of chitosan using chitinases or chitosanases, as the substrate specificity of the enzyme determines the composition of the oligomeric products. Protein‐engineering of these enzymes to alter their substrate specificity can overcome the limitations associated with naturally occurring enzymes and expand the spectrum of specific paCOS that can be produced. Here, engineering the substrate specificity of Bacillus sp. MN chitosanase is described for the first time. Two muteins with active site substitutions can accept N‐acetyl‐D‐glucosamine units at their subsite (?2), which is impossible for the wildtype enzyme. 相似文献
65.
Sarah M. Prexler Ratna Singh Bruno M. Moerschbacher Mareike E. Dirks-Hofmeister 《Plant molecular biology》2018,96(1-2):151-164
Key message
Successful site-directed mutagenesis combined with in silico modeling and docking studies for the first time offers experimental proof of the role of the ‘substrate selector’ residue in plant polyphenol oxidases.Abstract
The plant and fungi enzymes responsible for tissue browning are called polyphenol oxidases (PPOs). In plants, PPOs often occur as families of isoenzymes which are differentially expressed, but little is known about their physiological roles or natural substrates. In a recent study that explored these structure–function relationships, the eleven known dandelion (Taraxacum officinale) PPOs were shown to separate into two different phylogenetic groups differing in catalytic cavity architecture, kinetic parameters, and substrate range. The same study proposed that the PPOs’ substrate specificity is controlled by one specific amino acid residue positioned at the entrance to the catalytic site: whereas group 1 dandelion PPOs possess a hydrophobic isoleucine (I) at position HB2+1, group 2 PPOs exhibit a larger, positively charged arginine (R). However, this suggestion was only based on bioinformatic analyses, not experiments. To experimentally investigate this hypothesis, we converted group 1 ToPPO-2 and group 2 ToPPO-6 into PPO-2-I244R and PPO-6-R254I, respectively, and expressed them in E. coli. By performing detailed kinetic characterization and in silico docking studies, we found that replacing this single amino acid significantly changed the PPO’s substrate specificity. Our findings therefore proof the role of the ‘substrate selector’ in plant PPOs.66.
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Two dimensional metal nanostructures such as silver nanorods (AgNRs) exhibit a unique surface plasmon resonance (SPR) pattern involving transverse (TM) and longitudinal modes (LM). Such plasmonic near field is observed to have a profound effect on YAG:Ce nanoparticles placed in optimal proximity resulting in an unprecedented blue emission at TM excitation (383 nm). Unembellished YAG:Ce does not fluoresce under UV light incidence which implies that the phenomenon is exclusively due to modification of Ce3+ energy levels by plasmonic near field generated by AgNRs. Finite difference time domain (FDTD) electrodynamics simulation of near fields generated by exact AgNR and particle hybrids at TM, LM, and emission frequencies establish a cause-effect relationship. Plasmon-induced new emission from industrially important YAG:Ce nanophosphor opens up newer vistas for solid state lighting. 相似文献
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The role of circadian clocks in timing daily behaviors is widely acknowledged, and while empirical evidence suggests that clock period is correlated with the preferred phase of a rhythmic behavior (chronotype), other clock properties have also been hypothesized to underlie chronotype variation. Here, we report that fruit fly Drosophila melanogaster populations exhibiting evening emergence chronotype (late) are characterized by higher incidence of behavioral arrhythmicity in constant dim light, wider range of entrainment, reduced rates of re-entrainment to simulated jet-lag and higher amplitude of both entrained and free-running rhythms as compared to those exhibiting morning emergence chronotype (early). Our results thus highlight the role of circadian clock properties such as zeitgeber sensitivity, amplitude and coupling in driving chronotype variation. 相似文献
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Bijal Vyas Ratna D. Puri Narayanan Namboodiri Renu Saxena Mohan Nair Prahlad Balakrishnan M.P. Jayakrishnan Ameya Udyavar Ravi Kishore Ishwar C. Verma 《Indian pacing and electrophysiology journal》2016,16(1):8-18
BackgroundLong QT syndromes (LQTS) are characterized by prolonged QTc interval on electrocardiogram (ECG) and manifest with syncope, seizures or sudden cardiac death. Long QT 1–3 constitute about 75% of all inherited LQTS. We classified a cohort of Indian patients for the common LQTS based on T wave morphology and triggering factors to prioritize the gene to be tested. We sought to identify the causative mutations and mutation spectrum, perform genotype-phenotype correlation and screen family members.MethodsThirty patients who fulfilled the criteria were enrolled. The most probable candidate gene among KCNQ1, KCNH2 and SCN5A were sequenced.ResultsOf the 30 patients, 22 were classified at LQT1, two as LQT2 and six as LQT3. Mutations in KCNQ1 were identified in 17 (77%) of 22 LQT1 patients, KCNH2 mutation in one of two LQT2 and SCN5A mutations in two of six LQT3 patients. We correlated the presence of the specific ECG morphology in all mutation positive cases. Eight mutations in KCNQ1 and one in SCN5A were novel and predicted to be pathogenic by in-silico analysis. Of all parents with heterozygous mutations, 24 (92%) of 26 were asymptomatic. Ten available siblings of nine probands were screened and three were homozygous and symptomatic, five heterozygous and asymptomatic.ConclusionsThis study in a cohort of Asian Indian patients highlights the mutation spectrum of common Long QT syndromes. The clinical utility for prevention of unexplained sudden cardiac deaths is an important sequel to identification of the mutation in at-risk family members. 相似文献
70.
S Srivastava R S Phadke G Govil 《Physiological chemistry and physics and medical NMR》1990,22(2):105-113
Conformation of the renin inhibitor peptide, Pro-His-Pro-Phe-His-Phe-Phe-Val-Tyr-Lys (RIP) has been studied in aqueous solution and in lipid bilayers using 500 MHz 1H NMR spectroscopy. Analysis of the NMR parameters indicates that in aqueous solution, RIP exists as a random coil. On incorporation into lipid bilayers, the peptide adopts a rigid and well defined conformation. The N-terminal end is stabilized by the hydrophobic environment of the lipid bilayer. The C-terminal end is located near the lipid-water interface and attains rigidity due to interaction with the phosphate groups of lipids. The observations emphasize the role of environment in stabilizing significantly different conformations of RIP in three different media--D2O, DMSO and lipid bilayers. 相似文献