46 research outputs found
Chemoselective Installation of Amine Bonds on Proteins through Aza-Michael Ligation.
Chemical modification of proteins is essential for a variety of important diagnostic and therapeutic applications. Many strategies developed to date lack chemo- and regioselectivity as well as result in non-native linkages that may suffer from instability in vivo and adversely affect the protein's structure and function. We describe here the reaction of N-nucleophiles with the amino acid dehydroalanine (Dha) in a protein context. When Dha is chemically installed in proteins, the addition of a wide-range N-nucleophiles enables the rapid formation of amine linkages (secondary and tertiary) in a chemoselective manner under mild, biocompatible conditions. These new linkages are stable at a wide range of pH values (pH 2.8 to 12.8), under reducing conditions (biological thiols such as glutathione) and in human plasma. This method is demonstrated for three proteins and is shown to be fully compatible with disulfide bridges, as evidenced by the selective modification of recombinant albumin that displays 17 structurally relevant disulfides. The practicability and utility of our approach is further demonstrated by the construction of a chemically modified C2A domain of Synaptotagmin-I protein that retains its ability to preferentially bind to apoptotic cells at a level comparable to the native protein. Importantly, the method was useful for building a homogeneous antibody-drug conjugate with a precise drug-to-antibody ratio of 2. The kinase inhibitor crizotinib was directly conjugated to Dha through its piperidine motif, and its antibody-mediated intracellular delivery results in 10-fold improvement of its cancer cell-killing efficacy. The simplicity and exquisite site-selectivity of the aza-Michael ligation described herein allows the construction of stable secondary and tertiary amine-linked protein conjugates without affecting the structure and function of biologically relevant proteins
cDNA and Derived Amino Acid Sequence of the Chloroplastic Copper/Zinc-Superoxide Dismutase from Aspen (Populus tremuloides)
Cloning and Characterization of Antioxidant Genes From Populus Tremuloides (L.) Michx (Oxidative Stress, Superoxide Dismutase)
Genomic and cDNA clones, corresponding to an ozone-induced cytosolic copper–zinc superoxide dismutase, were isolated from quaking aspen (Populus tremuloides Michx.). The cytosolic superoxide dismutase (SOD) appears to be part of a multi-gene family in aspen and is interrupted by five introns in the coding region. Northern blot analysis with a gene-specific probe revealed an increase in the expression of this gene in response to ozone in the leaves of an ozone-tolerant aspen clone, compared with an ozone-sensitive clone. Cytosolic SOD transcript expression levels in leaves were also found to increase significantly within 6 h of mechanical wounding, after which the level of the transcript decreases. Under normal growing conditions, immature male and female aspen floral bud tissues contained the highest levels of the cytosolic SOD gene transcript, whereas transcript levels were almost undetectable in older leaves. © 1999 Elsevier Science Ireland Ltd. All rights reserved
cDNA and derived amino acid sequence of the chloroplastic copper/zinc-superoxide dismutase from aspen (Populus tremuloides).
Gene structure and expression of the aspen cytosolic copper/zinc-superoxide dismutase (PtSodCc1)
Genomic and cDNA clones, corresponding to an ozone-induced cytosolic copper–zinc superoxide dismutase, were isolated from quaking aspen (Populus tremuloides Michx.). The cytosolic superoxide dismutase (SOD) appears to be part of a multi-gene family in aspen and is interrupted by five introns in the coding region. Northern blot analysis with a gene-specific probe revealed an increase in the expression of this gene in response to ozone in the leaves of an ozone-tolerant aspen clone, compared with an ozone-sensitive clone. Cytosolic SOD transcript expression levels in leaves were also found to increase significantly within 6hof mechanical wounding, after which the level of the transcript decreases. Under normal growing conditions, immature male and female aspen floral bud tissues contained the highest levels of the cytosolic SOD gene transcript, whereas transcript levels wer
Influence of Polyhydroxybutyrate Content on the Crystallization Behavior of Polyamide 6‐polyhydroxy‐butyrate Blends
Influência da adição e da modificação química de uma carga mineral nanoparticulada nas propriedades mecânicas e no envelhecimento térmico de compósitos poliuretano/sisal Influence of a nanoparticulate mineral filler addition and chemical modification of the mechanical properties and thermal aging of PU/Sisal composites
Este trabalho trata do efeito da hibridização do reforço, através da adição de uma carga mineral (bentonita) nanoparticulada, no desempenho mecânico de compósitos poliuretano/sisal contendo 25% em massa de fibras de sisal e moldados por compressão. As propriedades mecânicas, de tração e impacto, dos sistemas compósitos estudados foram avaliadas em função do teor (0-10% em massa) e de modificações químicas da carga mineral. Os efeitos do envelhecimento térmico nas propriedades tênseis (sigma, E, épsilon) de compósitos selecionados também foram investigados. O reforço mineral (Bentonita sódica Brasgel-PA) foi empregado em quatro formas, a saber: a) como fornecido (sem tratamento), b) tratado com ácido clorídrico (0,6N), c) modificado com cloreto de dodecil dimetil benzil amônio (Dodigen) e d) modificado com brometo de cetil trimetil amônio (Cetremide). Os resultados indicam que a incorporação da bentonita eleva as propriedades mecânicas dos compósitos e que melhores propriedades foram obtidas quando a bentonita foi tratada com o ácido clorídrico. O envelhecimento térmico em tempos curtos (até 4 dias) provocou elevação no módulo elástico e resistência na ruptura dos compósitos, o que foi atribuído à pós-cura da matriz. Em tempos longos (32 dias) a exposição térmica causou decréscimo nas propriedades tênseis (sigma, E, épsilon) dos compósitos investigados, o que foi atribuído à degradação oxidativa da matriz e dos reforços. O compósito mais resistente ao envelhecimento térmico foi o híbrido cuja carga mineral foi organofilizada com o sal Cetremide. Anáslies por DRX e MEV indicam que a estrutura dos compósitos híbridos é um misto de micro e nanocompósito.<br>This work deals with filler hybridization effects, by the addition of a nanoparticulate mineral filler (bentonite), on the mechanical performance of compression molded Polyurethane/sisal composites with 25 wt % fiber content. Composite tensile and impact properties were evaluated as a function of mineral filler content (0-10 wt %) and chemical modifications. Thermal aging effects onf tensile (sigma, E, epsilon) properties of selected composites were also ascertained. The mineral filler (Brasgel PA sodium bentonite) was employed in the following forms: a) as received; b) treated with a 0,6N HCl solution; c) chemically modified with dodecyl dimethyl benzyl ammonium chloride (Dodigen) and d) chemically modified with cetyl trimethyl ammonium bromide (Cetremide). Our results show that bentonite addition increases the mechanical properties of PU/sisal composites and that best overall mechanical performances was achieved with addition of the hydrochloric acid trated mineral filler. Thermal aging for short times (up to 4 days) led to small increases in composite's elastic modulus na tensile strengths, which was attributed to post-curing of the matrix. Long thermal exposure (32 days) led to decreases in composite tensile properties (sigma, E, epsilon), which was attributed to oxidative degradation of both: matrix and sisal fibers. The most thermally resistant composite was the hybrid (PU/sisal-bentonite) whose mineral filler was chemically modified with Cetremide. DRX and SEM data indicate the hybrids with organofilized bentonites to be composed of micro and nanocomposite structures