Protein proteolytic degradation

Protein G. This vitamin K-dependent glycoproteia serine protease zymogen is produced ia the Hver. It is an anticoagulant with species specificity (19—21). Proteia C is activated to Proteia by thrombomodulin, a proteia that resides on the surface of endothefial cells, plus thrombin ia the presence of calcium. In its active form, Proteia selectively iaactivates, by proteolytic degradation. Factors V, Va, VIII, and Villa. In this reaction the efficiency of Proteia is enhanced by complex formation with free Proteia S. la additioa, Proteia activates tissue plasminogen activator, which... 

P. fluorescens has well-developed mechanisms for the secretion of proteins into the periplasm which facilitates S—S bond formation and proper N-terminal processing. It also allows one to reduce the formation of inclusion bodies and, thus, the additional costs caused by refolding processes. Proteolytic degradation of the expressed protein is also low, and very high... 

In the quest to find other plants that are suitable as bioreactors, various monocoty-ledonous and dicotyledonous species have been tested. These include corn [16], rice and wheat [17], alfalfa [18], potato [19, 20], oilseed rape [21], pea [22], tomato [23] and soybean [24]. The major advantage of cereal crops is that recombinant proteins can be directed to accumulate in seeds, which are evolutionar specialized for storage and thus protect proteins from proteolytic degradation. Recombinant proteins are reported to remain stable in seeds for up to five months at room temperature [17] and for at least three years at refrigerator temperature without significant loss of activity [25]. In addition, the seed proteome is less complex than the leaf proteome, which makes purification quicker and more economical [26]. 

In order to make molecular farming commercially profitable, recombinant proteins must be produced at a sufficiently high yield and in an active form. It has become clear that, for high-level protein accumulation, the stability of transgene expression can be as important as the expression level itself. The quantity of protein is determined by the rate of protein synthesis, assembly as well as proteolytic degradation [83]. 

Proteolytic degradation of a protein is characterized by hydrolysis of one or more peptide (amide) bonds in the protein backbone, generally resulting in loss of biological activity. Hydrolysis is usually promoted by the presence of trace quantities of proteolytic enzymes, but can also be caused by some chemical influences. 

Proteins differ greatly in their intrinsic susceptibility to proteolytic attack. Resistance to proteolysis seems to be dependent upon higher levels of protein structure (i.e. secondary and tertiary structure), as tight packing often shields susceptible peptide bonds from attack. Denaturation thus renders proteins very susceptible to proteolytic degradation. 

A number of strategies may be adopted in order to minimize the likelihood of proteolytic degradation of the protein product, these include ... 

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