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      Identification and Molecular Characterization of a Chitin-Binding Protein from the Beet Webworm, Loxostege sticticalis L.

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          Abstract

          As the first crucial barrier in the midgut of insects, the peritrophic membrane (PM) plays an important role in preventing external invasion. PM proteins, as the major components of the PM, determine the structure and function of this membrane. A new PM protein, named LstiCBP, from the PM of Loxostege sticticalis larvae was identified using cDNA library screening. The full cDNA of LstiCBP is 2606 bp in length and contains a 2403 bp ORF that encodes an 808-amino acid preprotein with a 15-amino acid as signal peptide. The deduced protein sequence of the cDNA contains 8 cysteine-rich chitin-binding domains (CBDs). Recombinant LstiCBP was successfully expressed in BL21 cells using recombinant plasmid DNA and showed high chitin-binding activity. LstiCBP expression was detected in the midgut at both the transcriptional and translational levels; however, the biochemical and physiological functions of LstiCBP in L. sticticalis require further investigation.

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          Protein Identification and Analysis Tools on the ExPASy Server

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            Peritrophic matrix structure and function.

            Formed of proteins, glycoproteins, and chitin microfibrils in a proteoglycan matrix, the peritrophic matrix (PM) separates the food from the midgut epithelium in most but not all insects. A PM occurs in two forms. A type I PM is delaminated from the entire midgut epithelium and, in some cases, may only be formed in response to feeding and the type of meal ingested. A type II PM is produced by a specialized region of the anterior midgut called the cardia and forms a continuous sleeve (or sleeves) that is always present. As it is positioned between food and midgut epithelium, the PM plays key roles in the intestinal biology of the insect. The PM may protect the midgut epithelium from mechanical damage and insult from pathogens and toxins; it must act as a semipermeable membrane regulating passage of molecules between the different midgut compartments; and it may separate the midgut lumen into different, physiologically significant compartments.
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              New insights into peritrophic matrix synthesis, architecture, and function.

              The peritrophic matrix (PM) is a chitin and glycoprotein layer that lines the invertebrate midgut. Although structurally different, it is functionally similar to the mucous secretions of the vertebrate digestive tract. The PM is a physical barrier, protecting the midgut epithelium from abrasive food particles, digestive enzymes, and pathogens infectious per os. It is also a biochemical barrier, sequestering and, in some cases, inactivating ingested toxins. Finally, the PM compartmentalizes digestive processes, allowing for efficient nutrient acquisition and reuse of hydrolytic enzymes. The PM consists of an organized lattice of chitin fibrils held together by chitin binding proteins. Glycans fill the interstitial spaces, creating a molecular sieve, the properties of which are dependent on the immediate ion content and pH. In this review, we have integrated recent structural and functional information to create a holistic model for the PM. We also show how this information may generate novel technologies for use in insect pest management.
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                Author and article information

                Contributors
                Role: External Editor
                Journal
                Int J Mol Sci
                Int J Mol Sci
                ijms
                International Journal of Molecular Sciences
                MDPI
                1422-0067
                22 October 2014
                October 2014
                : 15
                : 10
                : 19147-19161
                Affiliations
                [1 ]State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China; E-Mails: jyin@ 123456ippcaas.cn (J.Y.); tinyys@ 123456126.com (S.Y.); kbli@ 123456ippcaas.cn (K.L.)
                [2 ]Plant Science and Technology College, Beijing University of Agriculture, Beijing 102206, China
                Author notes
                [* ]Authors to whom correspondence should be addressed; E-Mails: guoweibua@ 123456163.com (W.G.); yzcao@ 123456ippcaas.cn (Y.C.); Tel./Fax: +86-108-071-5483 (W.G.); +86-106-281-5619 (Y.C).
                Article
                ijms-15-19147
                10.3390/ijms151019147
                4227266
                25340980
                90c6d777-682c-4f03-a8b1-1c6bc25be1d6
                © 2014 by the authors; licensee MDPI, Basel, Switzerland.

                This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 29 August 2014
                : 26 September 2014
                : 13 October 2014
                Categories
                Article

                Molecular biology
                peritrophic membrane protein,loxostege sticticalis,chitin-binding protein,lsticbp

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