Worthington core enzyme -- application field of trypsin

Worthington Core enzymes include ： Collagenase , Deoxyribonuclease I, Elastase , Papain , Ribonuclease , Trypsin, etc . Let's take a look Worthington  Trypsin .

Worthington  Trypsin background ：

Trypsin is a pancreatic serine protease , It has substrate specificity based on positively charged lysine and arginine side chains （Brown  and  Wold 1973）. This enzyme is secreted by the pancreas , Participate in food protein digestion and other biological processes . Trypsin is a medium-sized globular protein , Trypsinogen is produced as an inactive zymogen （Chen et al. 2009）.

AI Meijie Worthington Trypsin features ：

Bovine pancreas expresses two forms of trypsin , The major cationic form and the minor anionic form . These protein sequences have  72%  Identity of , And their coding area has  78%  Identity of .

Each of these proteins is further processed into alternative forms . Catalytic trypsin contains a flexible enzyme “ Autolytic ring ”（ Residue  G145-V157）（Schroeder  and  Shaw 1968  and  Bartunik wait forsomeone 1989）, And in the ring  K148-S149  The dominant single chain form  B- The autolysis of trypsin leads to the formation of A- Trypsin . stay  K193-D194  Further autolysis of leads to  Psi- The formation of trypsin （Fehlhammer  and  Bode 1975）.

Both cationic and anionic trypsin proteins are expressed in the form of trypsinogen , have  15  A residue signal peptide  (M1-A15)  And a  8  A residue propeptide  (F16-K23). The three-dimensional folding of all known trypsin is highly conserved . Besides , The regions flanking the catalytic triad and the catalytic triad are highly conserved （Hartley 1970）.  

AI Meijie Worthington Trypsin works ：

Trypsinogen can be activated by removing terminal hexapeptide to produce single chain β- Trypsin . Subsequent limited autolysis produces other active forms with two or more peptide chains bound by disulfide bonds . The main form is... With two peptide chains α- Trypsin and with a single chain β-.α- and β- Trypsin showed different activity and thermal stability .

Other structural features include amino acids  185-193  Surface ring at , Although not in direct contact with the substrate , But it will affect the specificity . Stability requires high affinity  Ca2+  Binding sites , When it doesn't exist , Autolysis occurs . Autolytic ring （ Located in amino acids  143-151） It is very flexible in trypsin and trypsinogen . The cleavage at lysine produces a substance that retains some catalytic activity α form . The protein has six completely conserved disulfide bonds （Halfon  and  Craik 1998）.

AI Meijie Worthington  Trypsin Application ：

1. Tissue dissociation , Especially when combined with other enzymes such as collagenase and elastase

2. adopt “ Trypsin digestion ” Harvest cells

3. Mitochondrial separation

4. In vitro study of proteins

5. Remove monolayer cells from plastic and glass

6. Various hemagglutination procedures

7. Flow cytometry  DNA  Sample preparation for analysis

8. Trypsin map

9. Fingerprinting and sequencing

10. The environmental monitoring

11. Reduce cell density in tissue culture

12. Passage cells

13. Cleave the fusion protein

14. Glycopeptides are produced from purified glycoproteins

AI Meijie Worthington Trypsin references ：

1.Abita, J.  and  Lazdunski, M.： About carboxylates in chymotrypsin  A  The structure and function of ： Chymotrypsin 、 Comparison of trypsinogen and trypsin , Biochem Biophys Res Commun 35 , 707, 1969

2.Abuchowski, A.  and  Davis, F.： polyethylene glycol - Preparation and properties of trypsin adduct , Biochim Biophys Acta 578,41,1979

3.Alagiri, S.  and  Singh, T.： Bifunctional amylase / Stability and kinetics of trypsin inhibitors , Biochim Biophys Acta 1203 , 77, 1993

4.Anson, M.： Estimate pepsin with hemoglobin 、 Trypsin 、 Papain and cathepsin , J Gen Physiol 22 , 79, 1938