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Cacao

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Contributes toGO:0003796lysozyme activityPMID:19881499IMP: Inferred from Mutant Phenotype F
This annotation made on page: BPP21:LYS
By: Portillosc (group Team Trypto-pham) on 2017-04-17 10:42:38 CDT.



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Team Trypto-pham0




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Entry TypeChallenging User,GroupTime/DateChallenge ReasonPoints/Assessment
Public
Assessment		DanielRenfro2017-05-08 22:14:01 CDT

This annotation has been flagged because it has been edited since last assessment

Qualifier GO ID GO term name Reference Evidence Code with/from Aspect Notes Status
GO:0003796 lysozyme activity PMID:19881499 IMP: Inferred from Mutant Phenotype F Supplementary Figure 2. is a graph of results of an assay to determine lysozyme activity between purified full-length R^21 and truncated R^21. The truncated version has had the N-terminal SAR (signal anchor release) domain cleaved. Lyz^P1, a protein from phage P1 that had been previously determined to have lyzosyme activity, was included for comparison. The results found that the truncated version of R^21 did not produce any lysozyme activity, while the full-length version produced activity similar to Lyz^P1. The assay was done with the EnzChek® Lysozyme Assay which measures lysozyme activity on fluorescently-labeled Micrococcus lysodeikticus cell walls. When the fluorescently-labeled Micrococcus lysodeikticus cell walls are hydrolyzed, the fluorescein was released from the cell wall resulting in an increase in fluorescence. Trials with different concentrations of egg white lysozyme (EWL) (a standard for the EnzChek test) were run. The different shapes in the graph indicate EWL concentrations. Different line colors indicate which protein was being tested. In each trial, the full-length version of R^21 produced similar activity to Lyz^P1, while the truncated version of R^21 experienced almost total loss of function. These results support the idea that the SAR domain is necessary for and contributes to lysozyme activity in R^21. complete
CACAO 12587
on BPP21:LYS
Flagged
Public
Assessment		AAJohnson2017-05-08 13:40:41 CDT

I don't believe you need a qualifier.

I like the GO term because we looked up the lysozyme assay kit used for this figure and they claim it measures hydrolysis of "β-(1-4)-glucosidic linkages between N-acetylmuramic acid and N-acetyl-D-glucosamine residues present in the mucopolysaccharide cell well of a variety of microorganisms." We are trusting the kit.

I think IMP works because the natural SAR was deleted and replaced with a different cleavable secretory signal sequence: "For production of iR21, BL21(DE3) cells were transformed with pETpelBΦ R21 27-165"

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