Difference between revisions of "Auxiliary Activity Family 10"

Revision as of 04:48, 22 July 2013

This page is currently under construction. This means that the Responsible Curator has deemed that the page's content is not quite up to CAZypedia's standards for full public consumption. All information should be considered to be under revision and may be subject to major changes.

Author: ^^^Vincent Eijsink^^^ and ^^^Gustav Vaaje-Kolstad^^^
Responsible Curator: ^^^Vincent Eijsink^^^

Auxiliary Activity Family 10
Clan	none, structurally related to AA9
Mechanism	lytic oxidase
Active site residues	mononuclear copper ion
CAZy DB link
http://www.cazy.org/AA10.html

Substrate specificities

Beware! Very preliminary text! AA10s have been shown to cleave chitin (REFs) and cellulose (REFs) chains through an oxidative reaction mechanism. However, since a substantial amount of studies have been published on members of the AA10 family prior to identification of their enzymatic function, substrate binding data is more abundant than data showing enzyme activity data. It should be noted that AA10 modules often are found combined with additional modules, e.g. carbohydrate binding modules (CBMs) that may determine or aid binding of the catalytic AA10 module to its substrate (see further down for examples).

Before the proteins belonging to AA10 were identified as enzymes, they were generally known as chitin binding proteins (CBPs). The reason for this is that most characterized proteins had been identified in chitinolytic systems such as that of Serratia marcescens (REF), several Streptomyces species (REFs), ......, and show no other obvious function than just binding the substrate. Thus there exists several papers that have analyzed the binding preferences of AA10s.

Shortly after CBP21 from S. marcescens was shown to specifically cleave chitin chains [1], CelS2 from Streptomyces coelicolor (also known as ScAA10D) was shown to act specifically on cellulose by a apparently identical monooxygenase activity [2]. In contrast to CBP21, which is a single AA10 module that binds strongly to beta-chitin, CelS2 has a CBM2 attached to the C-terminal side of the AA10 module that specifies binding of the enzyme cellulose.

Please see these references for an essential introduction to the CAZy classification system: [3, 4].

Kinetics and Mechanism

Content is to be added here.

Catalytic Residues

Content is to be added here.

Three-dimensional structures

In 2005 the structure CBP21 from S. marcescens was solved and represents the first structure in the AA10 family 2BEM [5]. The CBP21 wild type structure has three molecules in the asymetric unit, which of only chain C show electron density for a metal bound in the metal binding motif (modeled as a sodium ion, but is probably a reduced copper ion with low occupancy). Later the same year the structure of the CBP21-Y54A mutant was solved (different crystal form and space group), showing two molecules in the asymetric unit with no trace of electron density for a metal ion bound in the active site 2BEN [6]. The second AA10 structure, one of two AA10 from Burkholderia pseudomallei 1710b (Uniprot ID: Q3JY22), was published in the PDB late in 2011 by Seattle Structural Genomics Center for Infectious Disease 3UAM. The structure contains five molecules in the asymetric unit that all have two amino acids from the signal peptide still attached to the N-terminus, most likely disrupting the active site. The third unique AA10 structure to be solved was GbpA from Vibrio cholerae O1 biovar El Tor str. N16961 2XWX [7]. GbpA is unique in the sense that it contains four discreet modules (the N-termainl AA10 module, two modules with unkown funtion and a C-terminal CBM5/12) in addition to the N-terminal AA10 module. The published structure of GbpA only lacks the C-termainal CBM5/12 and is thus the first multimodular AA10 structure to be published. Shortly after the release of the GbpA, the structure of EfCBM33A from Enterococcus faecalis 4A02 [8] was published. The structure of EfCBM33A was solved at very high resolution (0.95Å), but similar to the other structures solved, no metal ion was observed bound in the active site. In 2012 the solution structure of CBP21 wild type (apo-form) was solved by NMR 2LHS [9] and in spring 2013 the structure of the single AA10 harbored byBacillus amyloliquefaciens DSM7 (BaCBM33) was published [10]. The latter publication contained three structures; the apo-enzyme 2YOW and two structures containing a reduced copper ion bond to the active site 2YOY 2YOX, thus being the first AA10 structures with the copper ion (that is essential for activity) bound in the active site.

Family Firsts

First stereochemistry determination: Content is to be added here.
First catalytic nucleophile identification: Content is to be added here.
First general acid/base residue identification: Content is to be added here.
First 3-D structure: CBP21, the single AA10-type LPMO from the Gram negative bacterium Serratia marcescens. Entry in the protein data bank: [1]

References

All Medline abstracts: PubMed

@@ Line 31: / Line 31: @@
 Beware!  Very preliminary text!
-AA10s have been shown to cleave chitin (REFs) and cellulose (REFs) chains through an oxidative reaction mechanism. However, since a substantial amount of studies have been published on members of the AA10 family prior to identification of their enzymatic function, substrate binding data is more abundant than data showing enzyme activity data.
+AA10s have been shown to cleave chitin (REFs) and cellulose (REFs) chains through an oxidative reaction mechanism. However, since a substantial amount of studies have been published on members of the AA10 family prior to identification of their enzymatic function, substrate binding data is more abundant than data showing enzyme activity data. It should be noted that AA10 modules often are found combined with additional modules, e.g. carbohydrate binding modules (CBMs) that may determine or aid binding of the catalytic AA10 module to its substrate (see further down for examples).
-Before the proteins belonging to AA10 were identified as enzymes, they were known as chitin binding proteins (CBPs). The reason for this is that most characterized proteins had been identified in chitinolytic systems such as that of ''Serratia marcescens'' (REF), several ''Streptomyces'' species (REFs), ......, and show no other obvious function than just binding the substrate. Thus there exists several papers that have analyzed the binding preferences of AA10s.
+Before the proteins belonging to AA10 were identified as enzymes, they were generally known as chitin binding proteins (CBPs). The reason for this is that most characterized proteins had been identified in chitinolytic systems such as that of ''Serratia marcescens'' (REF), several ''Streptomyces'' species (REFs), ......, and show no other obvious function than just binding the substrate. Thus there exists several papers that have analyzed the binding preferences of AA10s.
 Shortly after CBP21 from S. marcescens was shown to specifically cleave chitin chains <cite>,Vaaje-Kolstad2010-3</cite>, CelS2 from Streptomyces coelicolor (also known as ScAA10D) was shown to act specifically on cellulose by a apparently identical monooxygenase activity <cite>Forsberg2011</cite>. In contrast to CBP21, which is a single AA10 module that binds strongly to beta-chitin, CelS2 has a CBM2 attached to the C-terminal side of the AA10 module that specifies binding of the enzyme cellulose.