Purification and characterization of a novel chymotrypsin inhibitor controlled by the chymotrypsin inhibitor A (Ict-A) gene from the larval hemolymph of the silkworm, Bombyx mori

In Vivoandin VitroInteractions of theBombyx moriChymotrypsin Inhibitor b1 withEscherichia coli

“…Fax: +81-92-624-1011; E-mail: fujii@agr.kyushu-u.ac.jp Abbreviations: AF, Alexa Fluor; APNE, N-acetyl-DL-phenylalanine -naphthyl ester; BmLBP, Bombyx mori LPS-binding protein; CI, chymotrysin inhibitor; GNBP, Gram negative bacteria binding protein; LB, Luria-Bertani; LPS, lipopolysaccharide; PAGE, polyacrylamide gel electrophoresis; PBS, phosphate buffer saline; SDS-PAGE, sodium dodecylsulfate PAGE previously reported. 10) Polyacrylamide gel electrophoresis (PAGE) and sodium dodecylsulfate PAGE (SDS-PAGE) on a 15% separating gel were done by the methods previously reported. 10,17) Chymotrypsin-activity staining of a PAGE gel was done with -chymotrypsin, APNE, and tetrazotized orthodiamisidine by the method previously reported.…”

“…10) Polyacrylamide gel electrophoresis (PAGE) and sodium dodecylsulfate PAGE (SDS-PAGE) on a 15% separating gel were done by the methods previously reported. 10,17) Chymotrypsin-activity staining of a PAGE gel was done with -chymotrypsin, APNE, and tetrazotized orthodiamisidine by the method previously reported.…”

“…19,21) Compared with these values, the binding of CI-b1 to LPS was assumed to be C in a 0.1 M sodium phosphate buffer (pH 7.4) containing -chymotrypsin (12.5 g), CI-b1 (2 g), and LPS (0-50 g) was measured by the method previously reported. 10) Activity is the average of three measurements and is expressed as a percentage of that detected in the absence of CI-b1. very weak.…”

“…8) Biochemical and genetic properties of CIs have been reported. [9][10][11][12][13][14] But there exists a quite insufficient knowledge of the physiological roles of CI. CI-b1 is a Kunitztype CI with a basic pI.…”

Chromosomal localization of silkworm (Bombyx mori) sericin gene 1 and chymotrypsin inhibitor 13 using fluorescence in situ hybridization

“…There are at least sixteen kinds of CIs in silkworm, which are controlled by the complex genes in three linkage groups with codominance [12,13] . Chymotrypsin inhibitor 13 (CI-13), an inhibitor with low molecular weight, which controls the expression of 7000-10000 low molecular weight protease inhibitors, can be found in all silkworms and is controlled by the Ict-A gene localized at the 23.7 site of the second linkage group [14] .…”