Trypsin inhibitors soybeans

FIGURE l.l Hydrophobic interaction and reversed-phase chromatography (HIC-RPC). Two-dimensional separation of proteins and alkylbenzenes in consecutive HIC and RPC modes. Column 100 X 8 mm i.d. HIC mobile phase, gradient decreasing from 1.7 to 0 mol/liter ammonium sulfate in 0.02 mol/liter phosphate buffer solution (pH 7) in 15 min. RPC mobile phase, 0.02 mol/liter phosphate buffer solution (pH 7) acetonitrile (65 35 vol/vol) flow rate, I ml/min UV detection 254 nm. Peaks (I) cytochrome c, (2) ribonuclease A, (3) conalbumin, (4) lysozyme, (5) soybean trypsin inhibitor, (6) benzene, (7) toluene, (8) ethylbenzene, (9) propylbenzene, (10) butylbenzene, and (II) amylbenzene. [Reprinted from J. M. J. Frechet (1996). Pore-size specific modification as an approach to a separation media for single-column, two-dimensional HPLC, Am. Lab. 28, 18, p. 31. Copyright 1996 by International Scientific Communications, Inc.. Shelton, CT.]... 

There is evidence that protease inhibitors selectively regulate the activity of specific digestive enzymes at the level of gene expression (Rosewicz et al., 1989). Specifically, soybean trypsin inhibitor increases secretion of proteases, including a form of trypsin that is resistant to inhibition but does not cause an increase in amylase secretion. Although the relationships between protease inhibitors and exocrine pancreatic secretion have received the most attention, pancreatic secretion is increased when potato fiber is added to the diet (Jacob et al., 2000), although the mechanism and signaling pathway have not been elucidated. 

ROSEWicz s, LEWIS L D, WANG X Y, LiDDLE R A, LOGSDON c D (1989) Pancreatic digestive enzyme gene expression effects of CCK and soybean trypsin inhibitor. dm J Physiol. 256 G733-8. 

The following suggested protocol was developed by Barb Olson at Thermo Fisher for the labeling of soybean trypsin inhibitor (STI) with its subsequent complexation with trypsin. Modifications to this procedure may have to be done for other proteins. 

Fig. 5. Effect of the flow rate on the separation efficiency. Separation of a protein mixture at six different flow rates (40,80,120,160,200 and 240 ml/min) normalized to the elution volume. Conditions Column 80 ml CIM DEAE Tube Monolithic Column Mobile phase buffer A 20 mM Tris-HCl buffer, pH 7.4 buffer B 20 mM Tris-HCl buffer + 1 M NaCl, pH 7.4 Gradient 0-100% buffer B in 200 ml Sample 2 mg/ml of myoglobin (peak 1), 6 mg/ml of conalbumin (peak 2) and 8 mg/ml of soybean trypsin inhibitor (peak 3) dissolved in buffer A Injection volume 1 ml Detection UV at 280 nm. (Reprinted with permission from Podgornik A, Barut M, Strancar A, Josic D, Koloini T (2000) Anal Chem 72 5693)...

Fig. 29. An assortment of/3 barrels, viewed down the barrel axis (a) staphylococcal nuclease, 5-stranded (b) soybean trypsin inhibitor, 6-stranded (c) chymotrypsin, 6-stranded (d) immunoglobulin (McPC603 CH1) constant domain, 7-stranded (e) Cu,Zn superoxide dismutase, 8-stranded (f) triosephosphate isomerase, 8-stranded (g) im-... 

A. Up-and-down /3 barrels Papain domain 2 Soybean trypsin inhibitor Catalase domain 1... 

Soybean trypsin inhibitor, papain dl, and rubredoxin have identical topologies six strands of + 1,+ 1,+ 1,. . . proceeding to the left around the barrel if the chain termini are at the bottom. However, handedness is not nearly as meaningful a property for up-and-down topologies as it is for Greek keys, since up-and-down handedness can change on addition or delection of a single strand. 

Hexokinase Pyruvate kinase Adenylate kinase Phosphoglycerate kinase Phosphofructokinase Protease inhibitors Pancreatic trypsin inhibitor Soybean trypsin inhibitor Streptomyces subtilisin inhibitor Nucleases... 

I Soybean trypsin inhibitor (Kunitz-type) PF00197... 

II Soybean trypsin inhibitor (Bowman-Birk-type) PF00228... 

Fig. 2.1. Semilogarithmic plot of molecular weight (Mr) of marker proteins vs relative mobility (Rf) of marker proteins in gels of different acrylamide concentrations %T. Proteins 1 aprotinin (6.5 kD) 2 lysozyme (14.5 kD) 3 soybean trypsin inhibitor (21.5 kD) 4 carbonic acid anhydrase (31 kD) 5 hen ovalbumin (45 kD) 6 bovine serum albumin (66 kD) 7 phosphorylase b (97.4 kD) 8 8-galactosidase (116 kD) 9 myosin (205 kD)...

Yamanishi, R., Yusa, I., Miyamoto, A., Sato, I., Bando, N., and Terao, J. (2003). Alum augments the experimental allergenicity of Kunitz-t rpe soybean trypsin inhibitor independent of the antigen-adsorption. /. Nutr. Sci. Vitaminol. (Tokyo) 49, 409 13. 

Ribonuclease A, soybean trypsin inhibitor, thaumatin, a-lactalbumin AOT/isooctane Solubilization [71]... 

SeppMa,U.,Majamaa,H., Turjanmaa,K., Helin, J., Reunala, T., Kalkkinen,N., Palosuo, T. (2001). Identification of four novel potato (Solanum tuberosum) allergens belonging to the family of soybean trypsin inhibitors. Allergy, 56,619-626. 

Blrk and Applebaum (41) have studied the adverse effects of soybean trypsin inhibitors on development and protease activity in Tribolium castaneum. In Sitophilus oryzae, high doses of soybean trypsin inhibitor caused adult mortality (42). The wound-induced accumulation of these inhibitors is discussed by C. A. Ryan elsewhere in this symposium. 

Melmed, R.N., Elaaser, A.A.A., and Holt, S J. (1976). Hypertrophy and hyperplasia of neonatal rat exocrine pancreas induced by orally administered soybean trypsin-inhibitor. Biochimica Et Biophysica Acta, 421, 280-288. 

Materials and Methods. Fully deuterated phycocyanin and protio phycocyanin from Ph. luridum were used. The method of purifying phycocyanin was identical to that used previously (15, 16). The purity of the phycocyanin preparations, the complete substitution of deuterium for hydrogen in the fully deuterated phycocyanin, and the reversibility of the aggregation phenomenon were ascertained as previously (4, 16). Purified bovine trypsin, soybean trypsin inhibitor, and bovine liver catalase were obtained from the Worthington Biochemical Corp., Freehold, N. J., and used without further purification. Bovine a -casein B was kindly supplied by Chien Ho of the University of Pittsburgh. 

Literature values of sedimentation coefficients trypsin and soybean trypsin inhibitor (17) catalase (18) a,-casein B (JO). 

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