User:Tommie Hata/Introduction to Protein Engineering-Subtilisin: Difference between revisions

From Proteopedia
Jump to navigation Jump to search
← Older edit
No edit summary
No edit summary
 
(27 intermediate revisions by the same user not shown)
Line 3: Line 3:
<applet load='2sic' size='400' frame='true' align='right' caption='PDB ID 2sic, Subtilisin BPN' scene='User:Tommie_Hata/Introduction_to_Protein_Engineering-Subtilisin/Default-whole/1'/>
<applet load='2sic' size='400' frame='true' align='right' caption='PDB ID 2sic, Subtilisin BPN' scene='User:Tommie_Hata/Introduction_to_Protein_Engineering-Subtilisin/Default-whole/1'/>


The structure to the right is subtilisin BPN' and streptomyces subtilisin inhibitor (PDB ID: [http://www.pdb.org/pdb/explore/explore.do?structureId=2SIC 2sic]).  The subtilisin enzyme is colored white while the ''Streptomyces'' subtilisin inhibitor is colored yellow.  By <scene name='User:Tommie_Hata/Introduction_to_Protein_Engineering-Subtilisin/Default/3'>minimizing the inhibitor to a fragment that is bound in the subtilisin active site</scene>, we can take a look at how the substrate binds the active site.  Enzyme-inhibitor structures are common in the Protein Data Bank (PDB)compared to enzyme-substrate structures.  This is because enzyme-substrate complexes are often transient by nature.  The inability to form a stable enzyme-substrate complex makes it difficult to grow a crystal for structural determination through X-ray crystallography.  In comparison, enzyme inhibitors bind their targets with much higher affinity (low dissociation constant) which makes the enzyme-inhibitor complex


This is why many solved enzyme complexes  
== '''Introduction''' ==
The structure to the right is subtilisin BPN' and streptomyces subtilisin inhibitor (PDB ID: [http://www.pdb.org/pdb/explore/explore.do?structureId=2SIC 2sic]).  The subtilisin enzyme is colored white while the ''Streptomyces'' subtilisin inhibitor is colored yellow.  By <scene name='User:Tommie_Hata/Introduction_to_Protein_Engineering-Subtilisin/Default/3'>minimizing the inhibitor to a fragment that is bound in the subtilisin active site</scene>, we can take a closer look at the active site.  Enzyme-inhibitor structures are common in the Protein Data Bank (PDB)compared to enzyme-substrate structures.  This is because enzyme-substrate complexes are often transient.  The inability to form a stable enzyme-substrate complex makes it difficult to grow a crystal for structural determination through X-ray crystallography.  In comparison, enzyme inhibitors bind their targets with much higher affinity (low dissociation constant) which makes the enzyme-inhibitor complex more favorable to crystallize for structure determination.
 
<scene name='User:Tommie_Hata/Introduction_to_Protein_Engineering-Subtilisin/Default/1'>Return to default view</scene>
 
== '''Function''' ==
Subtilisin is a serine protease.  The <scene name='User:Tommie_Hata/Introduction_to_Protein_Engineering-Subtilisin/Labeled_active_site/1'>catalytic triad in the active site</scene> is made up of aspartic acid (Asp32), histidine (His64), and serine (Ser221).  Subtilisins are the proteases used in many laundry and dishwashing detergents.  In 2002 alone, 900 tons of subtilisin was produced and used in the EU.  Subtilisin received the first US patent for an engineered protein in 1988. 
 
<scene name='User:Tommie_Hata/Introduction_to_Protein_Engineering-Subtilisin/Default/1'>Return to default view</scene>
 
<scene name='User:Tommie_Hata/Introduction_to_Protein_Engineering-Subtilisin/Akr-1a80-play/1'>test link</scene>
 
== '''Engineered features''' ==
Subtilisins already possess a very high natural stability, partially because they are extracelluar enzymes optimized to work outside of the cell.  Yet there are additional characteristics that have been engineered into industrial subtilisins for turn them into better products.
 
'''A calcium-independent thermostable subtilisin BPN mutant''' (PDB ID: [http://www.pdb.org/pdb/explore/explore.do?structureId=1GNV 1gnv])
 
<applet load='1GNV-mod.pdb' size='300' frame='true' align='right' caption='PDB ID 1gnv, A calcium-independent thermostable subtilisin BPN mutant' />
<applet load='2sic' size='300' frame='true' align='right' caption='PDB ID 2sic, Subtilisin BPN' scene='User:Tommie_Hata/Introduction_to_Protein_Engineering-Subtilisin/2sic-highlighted/1'/>
 
 
----


Subtilisin is a serine protease.  The <scene name='User:Tommie_Hata/Introduction_to_Protein_Engineering-Subtilisin/Labeled_active_site/1'>catalytic triad in the active site</scene> is made up of aspartic acid (Asp32), histidine (His64), and serine (Ser221).




Line 20: Line 39:




==Reference==
=='''Reference'''==
'''Structures'''
 
<ref group="xtra">PMID:1920411</ref><references group="xtra"/>
<ref group="xtra">PMID:1920411</ref><references group="xtra"/>
'''Features and industrial use'''
<ref group="xtra">PMID:15296930</ref><references group="xtra"/>
'''Engineering'''
<ref group="xtra">PMID:15296930</ref><references group="xtra"/>

Latest revision as of 16:10, 23 October 2009

This Protein Engineering module has been developed using material from Dr. Scott Banta's course, Protein Engineering (chemical engineering department at Columbia University).

PDB ID 2sic, Subtilisin BPN

Drag the structure with the mouse to rotate


IntroductionIntroduction

The structure to the right is subtilisin BPN' and streptomyces subtilisin inhibitor (PDB ID: 2sic). The subtilisin enzyme is colored white while the Streptomyces subtilisin inhibitor is colored yellow. By , we can take a closer look at the active site. Enzyme-inhibitor structures are common in the Protein Data Bank (PDB)compared to enzyme-substrate structures. This is because enzyme-substrate complexes are often transient. The inability to form a stable enzyme-substrate complex makes it difficult to grow a crystal for structural determination through X-ray crystallography. In comparison, enzyme inhibitors bind their targets with much higher affinity (low dissociation constant) which makes the enzyme-inhibitor complex more favorable to crystallize for structure determination.

FunctionFunction

Subtilisin is a serine protease. The is made up of aspartic acid (Asp32), histidine (His64), and serine (Ser221). Subtilisins are the proteases used in many laundry and dishwashing detergents. In 2002 alone, 900 tons of subtilisin was produced and used in the EU. Subtilisin received the first US patent for an engineered protein in 1988.

Engineered featuresEngineered features

Subtilisins already possess a very high natural stability, partially because they are extracelluar enzymes optimized to work outside of the cell. Yet there are additional characteristics that have been engineered into industrial subtilisins for turn them into better products.

A calcium-independent thermostable subtilisin BPN mutant (PDB ID: 1gnv)

PDB ID 1gnv, A calcium-independent thermostable subtilisin BPN mutant

Drag the structure with the mouse to rotate

PDB ID 2sic, Subtilisin BPN

Drag the structure with the mouse to rotate



, which has been minimized in the view to highlight the chain bound in the active site of subtilisin.


ReferenceReference

Structures

[xtra 1]

  1. ↑ Takeuchi Y, Satow Y, Nakamura KT, Mitsui Y. Refined crystal structure of the complex of subtilisin BPN' and Streptomyces subtilisin inhibitor at 1.8 A resolution. J Mol Biol. 1991 Sep 5;221(1):309-25. PMID:1920411

Features and industrial use

[xtra 1]

  1. ↑ Maurer KH. Detergent proteases. Curr Opin Biotechnol. 2004 Aug;15(4):330-4. PMID:15296930 doi:10.1016/j.copbio.2004.06.005

Engineering

[xtra 1]

  1. ↑ Maurer KH. Detergent proteases. Curr Opin Biotechnol. 2004 Aug;15(4):330-4. PMID:15296930 doi:10.1016/j.copbio.2004.06.005
Retrieved from "https://proteopedia.openfox.io/wiki/index.php?title=User:Tommie_Hata/Introduction_to_Protein_Engineering-Subtilisin&oldid=1009692"