Nonribosomal peptide synthetases (NRPS), fatty acid synthases (FAS), and polyketide synthases (PKS) are multimodular enzymatic assembly lines utilized in natural product biosynthesis. The oligomeric structure of these assembly line enzymes has been a topic of interest because higher order oligomeric quaternary structural arrangements allow for alternate paths of acyl intermediate elongation and present unique challenges for the chimeric engineering of hybrid assembly lines. Unlike other NRPS systems that in general appear to be monomeric, the six domain (Cy1-Cy2-A-C1-PCP-C2) VibF subunit of vibriobactin synthetase has previously been shown to be dimeric, the same oligomeric state as that observed for FAS and PKS assembly lines. It has been demonstrated that the C1 domain within VibF is catalytically inactive and is not required for vibriobactin production. Utilizing sedimentation equilibrium analytical ultracentrifugation experiments to determine the oligomeric states of several VibF subfragments, we report that the C1 domain is largely responsible for VibF dimerization. Comparative rates of vibriobactin production, coupled with dissociation constants for VibF subfragment pair heterocomplexes, suggest that the mere presence of C1 does not detectably enhance the catalytic rates of neighboring domains, but it may properly orient Cy1-Cy2-A relative to PCP-C2. 相似文献
Engineered polyketide synthases (PKSs) are promising synthetic biology platforms for the production of chemicals with diverse applications. The dehydratase (DH) domain within modular type I PKSs generates an α,β-unsaturated bond in nascent polyketide intermediates through a dehydration reaction. Several crystal structures of DH domains have been solved, providing important structural insights into substrate selection and dehydration. Here, we present two DH domain structures from two chemically diverse PKSs. The first DH domain, isolated from the third module in the borrelidin PKS, is specific towards a trans-cyclopentane-carboxylate-containing polyketide substrate. The second DH domain, isolated from the first module in the fluvirucin B1 PKS, accepts an amide-containing polyketide intermediate. Sequence-structure analysis of these domains, in addition to previously published DH structures, display many significant similarities and key differences pertaining to substrate selection. The two major differences between BorA DH M3, FluA DH M1 and other DH domains are found in regions of unmodeled residues or residues containing high B-factors. These two regions are located between α3–β11 and β7–α2. From the catalytic Asp located in α3 to a conserved Pro in β11, the residues between them form part of the bottom of the substrate-binding cavity responsible for binding to acyl-ACP intermediates.
Parkinson's disease (PD) is the second most common neurodegenerative disorder. As there is no definitive diagnostic test,
its diagnosis is based on clinical criteria. Recently transcranial duplex scanning (TCD) of the substantia nigra in the brainstem
has been proposed as an instrument to diagnose PD. We and others have found that TCD scanning of substantia nigra duplex is
a relatively accurate diagnostic instrument in patients with parkinsonian symptoms. However, all studies on TCD so far have
involved well-defined, later-stage PD patients, which will obviously lead to an overestimate of the diagnostic accuracy of
TCD. 相似文献
This study demonstrates the use of bootstrap methods to estimate the total population of urban and periurban areas using satellite imagery and limited survey data. We conducted complete household surveys in 20 neighborhoods in the city of Bo, Sierra Leone, which collectively were home to 25,954 persons living in 1,979 residential structures. For five of those twenty sections, we quantized the rooftop areas of structures extracted from satellite images. We used bootstrap statistical methods to estimate the total population of the pooled sections, including the associated uncertainty intervals, as a function of sample size. Evaluations based either on rooftop area per person or on the mean number of occupants per residence both converged on the true population size. We demonstrate with this simulation that demographic surveys of a relatively small proportion of residences can provide a foundation for accurately estimating the total population in conjunction with aerial photographs. 相似文献
Synthetic Biology Open Language (SBOL) Visual is a graphical standard for genetic engineering. It consists of symbols representing DNA subsequences, including regulatory elements and DNA assembly features. These symbols can be used to draw illustrations for communication and instruction, and as image assets for computer-aided design. SBOL Visual is a community standard, freely available for personal, academic, and commercial use (Creative Commons CC0 license). We provide prototypical symbol images that have been used in scientific publications and software tools. We encourage users to use and modify them freely, and to join the SBOL Visual community: http://www.sbolstandard.org/visual. 相似文献