A new lectin was purified from tubers of Arum maculatum 1. by affinity chromatography on immobilized asialofetuin. Although this lectin is also retained on mannose-Sepharose 48, under the appropriate conditions free mannose is a poor inhibitor of its agglutination activity. Pure preparations of the Arum lectin apparently yielded a single polypeptide band of approximately 12 kD upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis. However, N-terminal sequencing of the purified protein combined with molecular cloning of the lectin have shown that the lectin is composed of two different 12-kD lectin subunits that are synthesized on a single large precursor translated from an mRNA of approximately 1400 nucleotides. Lectins with similar properties were also isolated from the Araceae species Colocasia esculenta (L.) Schott, Xanthosoma sagittifolium (L.) Schott, and Dieffenbachia sequina Schott. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration of the different Araceae lectins have shown that they are tetrameric proteins composed of lectin subunits of 12 to 14 kD. Interestingly, these lectins are the most prominent proteins in the tuber tissue. Evidence is presented that a previously described major storage protein of Colocasia tubers corresponds to the lectin.
A hydrostatic pressure of 1 .5 GPa induces changes in the secondary structure of bovine pancreatic tryspin inhibitor (BPTI) as revealed by the analysis of the amide I' band with Fourier-transform infrared (FTIR) spectroscopy in the diamond anvil cell. The features of the secondary structure remain distinct at high pressure suggesting that the protein does not unfold. The fitted percentages of the secondary structure elements during compression and decompression strongly suggest that the pressure-induced changes are reversible. The pressure-induced changes in the tyrosine side chain band are also reversible. The results demonstrate that the infrared technique explores different aspects of the behaviour of proteins in comparison with two published molecular dynamics studies performed up to 1
This study reports the characterization of the recombinant 7-kDa protein P2 from Sulfolobus solfataricus and the mutants F31A and F31Y with respect to temperature and pressure stability. As observed in the NMR, FTIR, and CD spectra, wild-type protein and mutants showed substantially similar structures under ambient conditions. However, midpoint transition temperatures of the denaturation process were 361, 334, and 347 K for wild type, F31A, and F31Y mutants, respectively: thus, alanine substitution of phenylalanine destabilized the protein by as much as 27 K. Midpoint transition pressures for wild type and F31Y mutant could not be accurately determined because they lay either beyond (wild type) or close to (F31Y) 14 kbar, a pressure at which water undergoes a phase transition. However, a midpoint transition pressure of 4 kbar could be determined for the F31A mutant, implying a shift in transition of at least 10 kbar. The pressure-induced denaturation was fully reversible; in contrast, thermal denaturation of wild type and mutants was only partially reversible. To our knowledge, both the pressure resistance of protein P2 and the dramatic pressure and temperature destabilization of the F31A mutant are unprecedented. These properties may be largely accounted for by the role of an aromatic cluster where Phe31 is found at the core, because interactions among aromatics are believed to be almost pressure insensitive; furthermore, the alanine substitution of phenylalanine should create a cavity with increased compressibility and flexibility, which also involves an impaired pressure and temperature resistance.
Aims. We selected a large sample of O-B stars that were considered as (candidate) slowly pulsating B, β Cep, and Maia stars after the analysis of their data. We analysed our new seven passband data collected for these stars during the first three years of scientific operations of the telescope. We performed a frequency analysis for 28 targets with more than 50 high-quality measurements to improve their variability classification. For the pulsating stars, we tried both to identify the modes and to search for rotationally split modes.Methods. We searched for frequencies in all the passbands and colours by using two independent frequency analysis methods and we applied a 3.6 S/N-level criterion to locate the significant peaks in the periodograms. The modes were identified by applying the method of photometric amplitudes for which we calculated a large, homogeneous grid of equilibrium models to perform a pulsational stability analysis. When both the radius and the projected rotational velocity of an object are known, we determined a lower limit for the rotation frequency to estimate the expected frequency spacings in rotationally split pulsation modes. Results. We detected 61 frequencies, among which 33 are new. We classified 21 objects as pulsating variables (7 new confirmed pulsating stars, including 2 hybrid β Cep/SPB stars), 6 as non-pulsating variables (binaries or spotted stars), and 1 as photometrically constant. All the Maia candidates were reclassified into other variability classes. We performed mode identification for the pulsating variables for the first time. The most probable value is 0, 1, 2, and 4 for 1, 31, 9, and 5 modes, respectively, including only 4 unambiguous identifications. For 7 stars we cannot rule out that some of the observed frequencies belong to the same rotationally split mode. For 4 targets we may begin to resolve close frequency multiplets.
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