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Valegh Faid Nouha Denguir Violaine Chapuis Nicolas Bihoreau Guillaume Chevreux 《Proteomics》2014,14(21-22):2460-2470
Human factor XI (hFXI) is a 160‐kDa disulphide‐linked homodimer zymogen involved in the coagulation cascade. Its deficiency results in bleeding diathesis referred to as hemophilia C. hFXI bears five N‐glycosylation consensus sites per monomer, N72, N108, N335 on the heavy chain and N432, N473 on the light chain. This study reports the first in‐depth glycosylation analysis of hFXI based on advanced MS approaches. Hydrophilic interaction LC and MS characterization and quantification of the N‐glycans showed that the two major forms are complex biantennary mono‐α2,6‐sialylated (A2S1, 20%) and bis‐α2,6‐sialylated structures (A2S2, 66%). Minor triantennary structures (A3S3F, ~1.5%; A3S3, ~2%) were also identified. MS analyses of intact hFXI revealed full occupation of two of the three heavy‐chain glycosites and almost full‐site occupancy of the light chain. Analysis of hFXI glycopeptides by LC‐MS/MS enabled site‐specific glycan profiling and occupancy. It was evidenced that N335 was not glycosylated and that N72 and N108 were fully occupied, whereas N432 and N473 were occupied at about 92 and 95%, respectively. We also identified a new glycosite of the noncanonical format NXC at N145, occupied at around 5%. These data provide valuable structural information useful to understand the potential roles of N‐glycosylation on hFXI function and could serve as a structural reference. 相似文献
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Micheline El Khoury Hichem Azzouz Alain Chavanieu Nouha Abdelmalak Joël Chopineau Mireille Kallassy Awad 《Archives of microbiology》2014,196(6):435-444
The aim of this study was to characterize new Bacillus thuringiensis strains that have a potent insecticidal activity against Ephestia kuehniella larvae. Strains harboring cry1A genes were tested for their toxicity, and the Lip strain showed a higher insecticidal activity compared to that of the reference strain HD1 (LC50 of Lip and HD1 were 33.27 and 128.61 μg toxin/g semolina, respectively). B. thuringiensis Lip harbors and expresses cry1Aa, cry1Ab, cry1Ac, cry1Ad and cry2A. DNA sequencing revealed several polymorphisms in Lip Cry1Aa and Cry1Ac compared to the corresponding proteins of HD1. The activation process using Ephestia kuehniella midgut juice showed that Lip Cry1A proteins were more stable in the presence of larval proteases. Moreover, LipCry1A proteins exhibited higher insecticidal activity against these larvae. These results indicate that Lip is an interesting strain that could be used as an alternative to the worldwide used strain HD1. 相似文献
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