Documente Academic
Documente Profesional
Documente Cultură
Jump to: navigation, search This article needs additional citations for verification. Please help improve this article by adding citations to reliable sources. Unsourced material may be challenged and removed. (March 2009)
Serine Peptidase
Crystal structure of bovine chymotrypsin. The catalytic residues are shown as yellow sticks. Rendered from PDB 1CBW.
Identifiers
Symbol
Ser
Crystal structure of Trypsin, a typical serine protease. Serine proteases (or serine endopeptidases) are enzymes that cleave peptide bonds in proteins, in which serine serves as the nucleophilic amino acid at the (enzyme's) active site.[1] They are
found ubiquitously in both eukaryotes and prokaryotes. Serine proteases fall into two broad categories based on their structure: chymotrypsin-like (trypsin-like) or subtilisin-like.[2] In humans, they are responsible for co-ordinating various physiological functions, including digestion, immune response, blood coagulation and reproduction.[1]
Contents
[hide]
1 Chymotrypsin-like o 1.1 Trypsin-like o 1.2 Chymotrypsin-like o 1.3 Elastase-like 2 Subtilisin-like 3 Catalytic Mechanism o 3.1 Additional stabilizing effects 4 Regulation of Serine Protease Activity o 4.1 Zymogen Activation o 4.2 Inhibition 5 Role in disease 6 Diagnostic use 7 References 8 See also 9 External links
[edit] Chymotrypsin-like
Chymotrypsin-like serine proteases are characterised by a distinctive structure, consisting of two beta-barrel domains that converge at the catalytic active site. These enzymes can be further categorised based on their substrate specificity as either trypsin-like, chymotrypsin-like or elastase-like.[3]
[edit] Trypsin-like
Trypsin-like proteases cleave peptide bonds following a positively charged amino acid (lysine or arginine).[4] This specificity is driven by the residue which lies at the base of the enzyme's S1 pocket (generally a negatively charged aspartic acid or glutamic acid).
[edit] Chymotrypsin-like
The S1 pocket of chymotrypsin-like enzymes is more hydrophobic than in trypsin-like proteases. This results in a specificity for medium to large sized hydrophobic residues, such as tyrosine, phenylalanine and leucine.
[edit] Elastase-like
Elastase-like proteases have a much smaller S1 cleft than either trypsin- or chymotrypsin-like proteases. Consequently, residues such as alanine, glycine and valine tend to be preferred.
[edit] Subtilisin-like
Subtilisin is a serine protease in prokaryotes. Subtilisin is evolutionary unrelated to the chymotrypsin-clan, but shares the same catalytic mechanism utilising a catalytic triad, to create a nucleophilic serine. This is the classic example used to illustrate convergent evolution, since the same mechanism evolved twice independently during evolution.
The serine has an -OH group that is able to act as a nucleophile, attacking the carbonyl carbon of the scissile peptide bond of the substrate. A pair of electrons on the histidine nitrogen has the ability to accept the hydrogen from the serine -OH group, thus coordinating the attack of the peptide bond. The carboxyl group on the aspartic acid in turn hydrogen bonds with the histidine, making the nitrogen atom mentioned above much more electronegative.
The polypeptide substrate binds to the surface of the serine protease enzyme such that the scissile bond is inserted into the active site of the enzyme, with the carbonyl carbon of this bond positioned near the nucleophilic serine. The serine -OH attacks the carbonyl carbon, and the nitrogen of the histidine accepts the hydrogen from the -OH of the [serine] and a pair of electrons from the double bond of the carbonyl oxygen moves to the oxygen. As a result, a tetrahedral intermediate is generated. The bond joining the nitrogen and the carbon in the peptide bond is now broken. The covalent electrons creating this bond move to attack the hydrogen of the histidine, breaking the connection. The electrons that previously moved from the carbonyl oxygen double bond move back from the negative oxygen to recreate the bond, generating an acyl-enzyme intermediate. Now, water comes in to the reaction. Water replaces the N-terminus of the cleaved peptide, and attacks the carbonyl carbon. Once again, the electrons from the double bond move to the oxygen making it negative, as the bond between the oxygen of the water and the carbon is formed. This is coordinated by the nitrogen of the histidine, which accepts a proton from the water. Overall, this generates another tetrahedral intermediate.
In a final reaction, the bond formed in the first step between the serine and the carbonyl carbon moves to attack the hydrogen that the histidine just acquired. The now electrondeficient carbonyl carbon re-forms the double bond with the oxygen. As a result, the Cterminus of the peptide is now ejected.
Trypsinogen
trypsin
After the Arg 15 - Ile 16 bond in the chymotrypsinogen zymogen is cleaved by trypsin, the newly generated structure Chymotrypsinogen chymotrypsin called a pi-chymotrypsin undergoes autolysis (self digestion), yielding active chymotrypsin. Proelastase elastase It is activated by cleavage through trypsin. As can be seen, trypsinogen activation to trypsin is essential, because it activates its own reaction, as well as the reaction of both chymotrypsin and elastase. Therefore, it is essential that this activation does not occur prematurely. There are several protective measures taken by the organism to prevent self-digestion:
The activation of trypsinogen by trypsin is relatively slow The zymogens are stored in zymogen granules, capsules that have walls that are thought to be resistant to proteolysis.
[edit] Inhibition
There are certain inhibitors that resemble the tetrahedral intermediate, and thus fill up the active site, preventing the enzyme from working properly. Trypsin, a powerful digestive enzyme, is generated in the pancreas. Inhibitors prevent self-digestion of the pancreas itself. Serine proteases are paired with serine protease inhibitors, which turn off their activity when they are no longer needed.[5] Serine proteases are inhibited by a diverse group of inhibitors, including synthetic chemical inhibitors for research or therapeutic purposes, and also natural proteinaceous inhibitors. One family of natural inhibitors called "serpins" (abbreviated from serine protease inhibitors) can form a covalent bond with the serine protease, inhibiting its function. The best-studied serpins are antithrombin and alpha 1-antitrypsin, studied for their role in coagulation/thrombosis and emphysema/A1AT, respectively. Artificial irreversible small molecule inhibitors include AEBSF and PMSF.
Coagulation factor levels may be required in the diagnosis of hemorrhagic or thrombotic conditions.
Fecal elastase is employed to determine the exocrine activity of the pancreas, e.g., in cystic fibrosis or chronic pancreatitis. Serum prostate-specific antigen is used in prostate cancer screening, risk stratification, and post-treatment monitoring.
[edit] References
1. ^ a b Hedstrom, L. (Dec 2002). "Serine protease mechanism and specificity.". Chem Rev 102 (12): 450124. doi:10.1021/cr000033x. PMID 12475199. 2. ^ Madala PK, Tyndall JD, Nall T, Fairlie DP. (Jun 2010). "Update 1 of: Proteases universally recognize beta strands in their active sites". Chem Rev 110 (6): PR131. doi:10.1021/cr900368a. PMID 20377171. 3. ^ Ovaere P, Lippens S, Vandenabeele P, Declercq W. (Aug 2009). "The emerging roles of serine protease cascades in the epidermis". Trends Biochem Sci 34 (9): 45363. doi:10.1016/j.tibs.2009.08.001. PMID 19726197. 4. ^ Evnin, Luke B.; Vsquez, John R.; Craik, Charles S. (1990). "Substrate specificity of trypsin investigated by using a genetic selection". Proceedings of the National Academy of Sciences of the United States of America 87 (17): 665963. doi:10.1073/pnas.87.17.6659. JSTOR 2355359. PMC 54596. PMID 2204062. http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pmcentrez&artid=54596. 5. ^ Kimball's Biology Pages, Serine Proteases[self-published source?]
The MEROPS online database for peptidases and their inhibitors: Serine Peptidase Serine Proteases site at Saint Louis University (SLU) MeSH Serine+proteases [show]
v t e
3.4.13 Dipeptidase (1, 2, 3) 3.4.14 Dipeptidyl peptidase (Cathepsin C, Dipeptidyl peptidase-4) Tripeptidyl
peptidase (Tripeptidyl peptidase I, Tripeptidyl peptidase II) 3.4.15 Angiotensin-converting enzyme 3.4.16 Serine type carboxypeptidases: Cathepsin A DD-transpeptidase 3.4.17 Metallocarboxypeptidases: Carboxypeptidase (A, A2, B, C, E, Glutamate II)
Other/ungrouped Metalloexopeptidase
B enzm (
o o o o o o o o o o o o o
1.1 2 3 4 5 6 7 8 10 11 13 14 15-18 )
2.1 (
o o o o o o o
2 3 4 5 6 7 8
2 3 4 5 6 7 )
3.1.3.48 3.4.21 (
o o o
22 23 24 )
4.1 (
o o o o o
2 3 4 5 6
5.1 (
o o o o
2 3 4 99 )
6.1-3 (
o o
4 5-6 ) [show]
v t e
B enzm
(
o o o o o o o o o o o o o
1.1 2 3 4 5 6 7 8 10 11 13 14 15-18 )
2.1 (
o o o o o o o
2 3 4 5 6 7 8 )
2 3 4
o o o
5 6 7 )
3.1.3.48 3.4.21 (
o o o
22 23 24 )
4.1 (
o o o o o
2 3 4 5 6 )
5.1 (
o o o o
2 3 4 99
6.1-3 (
o o
4 5-6 )
Retrieved from "http://en.wikipedia.org/w/index.php?title=Serine_protease&oldid=484122525" Categories: EC 3.4.21 Hidden categories: Accuracy disputes from May 2011 Articles needing additional references from March 2009 All articles needing additional references Protein pages needing a picture
Personal tools
Namespaces
Article Talk
Navigation
Main page Contents Featured content Current events Random article Donate to Wikipedia
Interaction Toolbox
What links here Related changes Upload file Special pages Permanent link Cite this page
Print/export
Languages
Deutsch Espaol Italiano Portugus Svenska This page was last modified on 27 March 2012 at 03:37. Text is available under the Creative Commons Attribution-ShareAlike License; additional terms may apply. See Terms of use for details. Wikipedia is a registered trademark of the Wikimedia Foundation, Inc., a non-profit organization.