Insulin trypsin inhibitors

Materials. NIPAAm obtained from Fisher Scientific Inc. (Fair Lawn, NJ), was recrystallized from hexane. BMA and AA were purchased from Aldrich Chemical Company (Milwaukee, WI). BMA and AA were purified by vacuum distillation at 57 C / 17 mm Hg and 39 C / 10 mm Hg respectively. 2,2 -azobis-isobutyronitrile (AIBN), purchased from Eastman Kodak Company (Rochester, NY), was recrystallized firom methanol. Bovine insulin, trypsin inhibitor and angiotensin II were purchased fix>m Sigma Chemical Company (St Louis, MO). Human calcitonin was a gift from Suntory Ltd. (Tokyo, Japan). Heavy white mineral oil and decane were purchased fix>m Aldrich Chemical Company (Milwaukee, WI). Acetonitrile, HPLC grade, was purchased from Fisher Scientific Inc. (Fair Lawn, NJ). All other chemicals were reagent grade. 

Other SS-rich Pancreatic trypsin inhibitor Insulin... 

Abbreviations MBN, myoglobin MGN, myogen MHN, myohemerythrin RUB, mbredoxin ICG, immunoglobulin constant region IGV, immunoglobulin variable region PBN, prealbumin SDM, superoxide dismutase CON, conconavalin A CHT, chymotrypsin INS, insulin PTI, pancreatic trypsin inhibitor CB5, cytochrome b5 ... 

One of the hallmarks of OBPs is the six cysteine (six half cystines) residues, but this criterion alone is not sufficient to classify a certain protein as olfactory protein. It is important to demonstrate that an OBP is expressed only (or predominantly) in olfactory tissues. Evidence for their ability to bind odorants is also desirable, but not sine qua non. One of these criteria alone would not be enough to define a given protein as an OBP. For example, bovine serum albumin (BSA) binds to insect pheromones (Leal, unpublished data) and yet it is not an OBP because it does not occur in olfactory tissues in the first place. Conversely, a protein specific to antennae is not necessarily an OBP. There are other proteins that may be expressed in antennae but not in control tissues. Non-OBPs specific to insect antennae have been previously detected (Ishida and Leal, unpublished data). Also, a g lu tath i o n e -. S -1 ra n s I e ra s e has been reported to be expressed specifically in antennae of M. sexta (Rogers et al., 1999). Likewise, the six-cysteine criterion should not be misleadingly used. Insulin and bovine pancreatic trypsin inhibitor, for example, have six cysteines in three disulfide bridges and yet they are not odorant-binding proteins. Also, mammalian and insect defensins have six well-conserved cysteine residues. 

Coadministration of protease inhibitors can optimize protein absorption. The administration of soybean trypsin inhibitor improves die absorption of insulin... 

Oral dosage forms may contain various other additives to increase the solubility and hence oral bioavailability of the drag, such as co-solvents, buffers and surfactants. Newer technologies may also incorporate additives such as enzyme inhibitors, to prevent premature degradation of enzymatically labile drags. For example, the inclusion of trypsin inhibitors, such as soyabean trypsin inhibitor and aprotinin, have been shown to be effective in enhancing the effect of insulin in rats. Penetration enhancers may also be included to facilitate the uptake of poorly absorbed moieies. These are discussed below in Section 6.7.4. 

FIGURE 13 Plot of the logarithm of the retention volume (In VR) versus the concentration of the displacing salt, ammonium sulphate, in the HP-HIC mode with the proteins I, insulin B-chain 2, bovine trypsin inhibitor 3, bovine trypsinogen 4, insulin A-chain 5, ribonuclease 6, sperm whale myoglobin 7, horse heart cytochrome c. Data from Ref. 42. 

Carbonic anhydrase, a-lactalbumin, trypsin inhibitor, oval-bumn, conalbumin, hemoglobin variants Ribonuclease, insulin, a-lactalbumin Lysozyme, a-chymotrypsinogen, ribonuclease A, cytochrome c... 

Figure 2. SDS gel electrophoresis of the products of partial cystine cleavage for several test proteins. A. molecular weight standards, B. yeast alcohol dehydrogenase. C. P-lactoglobulin, D. hen egg lysozyme, E. ovalbumin, F. calf fetal serum fetuin. Molecular weight standards are indicated by arrows on the left side of the gel and are bovine serum albumin (66,300), bovine liver glutamate dehydrogenase (55,400), porcine muscle lactate ddiydiogenase (36,500), bovine erythrocyte carbonic anhydrase (31,000), soybean trypsin inhibitor (21,500), hen egg lysozyme (14,400), bovine lung aprotinin (6,000), unresolved bovine pancreatic insulin A and B chains.

Insulin Soybean trypsin inhibitor Chymostatin Aprotinin Bowman-Birk inhibitor Rat... 

Inbibin (activin), beta A Inhibin (activin), beta B Inbibin (activin), beta C Inhibin (activin), beta E Inhibin, alpha Insulin C-peptide Insulin, A chain Insulin, B chain Insulin-like growth factor lA Insulin-like growth factor II Inter-alpha trypsin inhibitor, HI Inter-alpha trypsin inhibitor, H2 Inter-alpha trypsin inhibitor, H4 Inter-alpha trypsin inhibitor, L Interferon alpha Interferon beta Interferon gamma Interleukin-1 beta Interleukin-10 Interleukin-12, alpha Interleukin-12, beta Interleukin-1 receptor antagonist Interleukin-2 Interleukin-4... 

Phe(J)B ]insulm Porcine proinsulin Acetamidino-insulin Bic(Boc) insulin Des-Al-amino-insulin Destripeptide B - bovine insulin 58-Unit Peptides Bovine basic pancreatic trypsin inhibitor... 

Wt of polypeptide drug, using insulin, calcitonin, trypsin inhibitor and angiotensin II as model polypeptides. Release rates fix>m these systems were controlled by varying the molecular weight (M. Wt.) of the polymers (19), and insulin was used as a model polypeptide drug. 

Fig. 3.13. Proportionality of the loss of accessible surface area to the molecular weight of proteins (from Janin, 1976). Different proteins insulin, rubredoxin, pancreatic trypsin inhibitor, HIPIP, calcium binding protein, ribonuclease S, lysozyme, staphylococcal nuclease, papain, chymotrypsin, concanavalin A, subtilisin, thermolysin, carboxypeptidase A.

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