Protease treatment

Roth, A. F., and Ward, W. W. (1983). Conformational stability after protease treatment in Aequorea GFP. Photochem. Photobiol. 37S S71. 

In vitro and ex vivo studies have shown that FATPs transport LCFAs and very long-chain fatty acids (VLCFAs) but no medium-chain fatty acids, fatty acid esters, or lipid-soluble vitamins [4]. LCFA transport is inhibited by prior protease treatment. Synthetic substrates for FATPs include 14C-labeled fatty acids and the fluorescently labeled fatty acid analogue C1 -BODEP Y-Cl 2. Using the latter substrate, differences in fatty acid uptake kinetics between FATP expressing 3T3 LI adipocytes and 3T3 LI fibroblasts, which are devoid of FATPs, can be readily appreciated (Fig. 2). 

Complete removal of degraded or damaged portions of the wool (not merely the cuticle) using (i) protease treatment (ii) formic acid rinse and application of a softener. 

Only empirical tests have so far been carried out, however [76]. In a detailed but small-scale study, various options were examined for the sequence (i) oxidative treatment (ii) protease treatment (iii) application of softener, including exhaust or pad application [133]. 

Antigenic determinants masked by formalin-fixation and paraffin-embedding may also be exposed by enzymatic digestion. This can, however, not be used with frozen sections or cells which are not paraffin-embedded. The beneficial effects of protease treatment are presumably related to cleavage of the molecular cross-links by the... 

Braun, Melsungen AG). The resulting cell-free suspension was subjected to protease treatment. This was accomplished by incubating the suspension with protease (2 mg/ml) for 4 hrs at 37°C with continuous shaking. 

Partial Purification. The cell-free suspension, before and after protease treatment, was subjected to gel-filtration chromatography on Sephadex G-75 according to the procedure described in our earlier report (li). Peak II of the sephadex column eluate will be referred to as flavoprotein preparation (Fig. 1). 

To study the effect of the protease treatment cell-free suspension, with or without protease treatment, was subjected to gel-filtration chromatography on Sephadex G-75 and the elution patterns were compared (Fig. 1). In each case, two major peaks were detected by monitoring column fractions with absorbance at 280 nm. Degradation activities on mexacarbate, in the presence of FMN and light under anaerobic condition, were measured for each fraction. It was found that the highest activity was associated with peak II. It is interesting to note that protein (s) associated with peak II were detected with or without protease treatment these will be referred to as natural flavoprotein (B, Fig. 

TX-20 cell-free preparation (A), after and (B), before protease treatment. (-),... 

The hydrophobic peptide segments of El and E2, which attach the spike protein to the lipid bilayer, can be localized on the polypeptide chains by a mapping procedure first used by Dintzis (1961) to show that the synthesis of polypeptide chains begins at the amino-terminal end. The hydrophobic stubs left in the viral membrane after protease treatment are found at the carboxyl-terminal ends of both the El and the E2 polypeptides (Garoff and Sdderlund, 1978). 

Protease treatment After washing, cells are exposed to a solution of 0.04% protease (pronase E) in Ca TMg free HEPES-containing ethylene glycol tetra acetic acid (EGTA) (Sigma) for 15 minutes at 37°C as applied in (93). Cells are then washed three times with ice-cold phosphate buffered saline (PBS) containing 0.1% BSA. 

Terminate the protease treatment by washing cells four times for 2 min each in PBS with gentle agitation (see Note 5). 

Nonspecific hybridization Does a noncomplementary probe of identical size and G-C composition hybridize Does a nuclease-treated specimen show hybridization Does protease treatment decrease hybridization ... 

Verma and McCalla ( ) studied the action of pepsin, papain and a commercial fungal protease on wheat gluten. All enzymes acted effectively on dispersed gluten however, the action of different enzymes produced different types of digestion products. Depending upon desired handling characteristics of bread doughs prepared from treated wheat flour, various types of protease treatments can be selected. 

DNA polymerase I, then, is not the primary enzyme of replication instead it performs a host of clean-up functions during replication, recombination, and repair. The polymerase s special functions are enhanced by its 5 —>3 exonuclease activity. This activity, distinct from the 3 —>5 proofreading exonuclease (Fig. 25-7), is located in a structural domain that can be separated from the enzyme by mild protease treatment. When the 5 —>3 exonuclease domain is removed, the remaining fragment (Afr 68,000), the large fragment or Klenow fragment (Fig. 25-8), retains the polymerization and... 

The quaternary structure of tyrosinase differs depending on the species. The tyrosinase can be found in either latent or active form, and the activating conditions differ depending on the species. Tyrosinase from the mushroom Agaricus bisporus is a heterotetramer that is compound of two subunits of 43 kDa (H) and two subunits of 13 kDa (L), with a native molecular mass of 111 kDa, and contains four copper atoms [152, 153], The isolated subunits do not possess the enzymatic activity. Activation of the mushroom pro-tyrosinase can be effected by protease treatment [154] or by SDS [155], Mushroom has an isozyme of... 

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