Identification and molecular characterization of a new member of the peritrophic membrane proteins from the meadow moth, loxostege sticticalis.
The peritrophic membrane (PM) plays an important role in protecting insects. The PM proteins are important to determinate the formation and function of the PM. A new PM protein, named Lsti99, was identified from the PM of Loxostege sticticalis larvae by cDNA library screening. The full cDNA of Lsti99 is 1392 bp in length, contains an open reading frame (ORF) of 1245 bp that encodes a preprotein of 415 amino acid residues with a 17-amino acid signal peptide. The sequence of Lsti99 showed no homology to other known PM proteins. The recombinant Lsti99 was successfully expressed in insect cells (Sf9) using recombinant baculoviruses and was used to isolate the antibodies to Lsti99 from the polyclonal antiserum. Lsti99 was expressed mainly in the PM, but weaker bands could be detected in the head and integument as well. The Lsti99 protein could be separated from the PM complex by chitinase in vitro, but M2R did not show effect in vitro confirming the chitin-binding activity of Lsti99. The biochemical and physiological functions of Lsti99 in L. sticticalis require further investigation.
Yin
Jiao
J
State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China.
Wei
Zhao-Jun
ZJ
Li
Ke-Bin
KB
Cao
Ya-Zhong
YZ
Guo
Wei
W
D000818:Animals; D001483:Base Sequence; D002688:Chitinases / Q000494:pharmacology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007814:Larva / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009036:Moths / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000032:analysis; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein
D018076:DNA, Complementary; D019476:Insect Proteins; D008565:Membrane Proteins; D011993:Recombinant Fusion Proteins; D002688:Chitinases
Biotech Histochem. 2003 Jun-Aug;78(3-4):201-5
14714884
Cell Tissue Res. 2005 Apr;320(1):175-85
15726420
Nucleic Acids Res. 2002 Jan 1;30(1):235-8
11752303
Glycoconj J. 1998 Feb;15(2):115-30
9557871
Insect Biochem Mol Biol. 2003 Feb;33(2):239-52
12535682
Insect Mol Biol. 2003 Aug;12(4):333-43
12864913
Arch Insect Biochem Physiol. 2001 Jun;47(2):62-75
11376453
Proc Natl Acad Sci U S A. 1997 Aug 19;94(17):8939-44
9256413
J Biol Chem. 1998 Jul 10;273(28):17665-70
9651363
J Insect Physiol. 2010 Sep;56(9):1325-31
20433846
J Insect Physiol. 2005 Jun;51(6):691-700
15869761
Insect Biochem Mol Biol. 1998 Feb;28(2):99-111
9639876
J Mol Biol. 1990 Oct 5;215(3):403-10
2231712
Arch Insect Biochem Physiol. 2001 Jun;47(2):110-8
11376457
Insect Biochem Mol Biol. 2000 Feb;30(2):135-43
10696589
Proc Natl Acad Sci U S A. 1997 Jun 24;94(13):6977-82
9192677
Insect Biochem Mol Biol. 2004 Oct;34(10):1101-15
15475304
Insect Biochem Mol Biol. 2004 Mar;34(3):215-27
14871618
Mol Biol Rep. 2008 Sep;35(3):329-35
17525867
Insect Biochem Mol Biol. 1999 Feb;29(2):87-101
10196732
Insect Biochem Mol Biol. 2005 Sep;35(9):947-59
15978997
Insect Biochem Mol Biol. 2005 Nov;35(11):1224-34
16203204
Insect Biochem Mol Biol. 2002 Dec;32(12):1663-71
12429118
J Mol Biol. 2004 Jul 16;340(4):783-95
15223320
Annu Rev Entomol. 1997;42:525-50
15012322
J Biol Chem. 2001 May 4;276(18):15527-36
11278497
J Biol Chem. 1996 Apr 12;271(15):8925-35
8621536
Insect Biochem Mol Biol. 2002 May;32(5):517-23
11891128
Insect Mol Biol. 2002 Jun;11(3):207-16
12000639
Insect Biochem Mol Biol. 2001 Jul 26;31(9):919-32
11439251
J Biol Chem. 1997 Jun 27;272(26):16663-9
9195982