Literature summary for 1.11.1.21 extracted from

Munir, A.; Wilson, M.T.; Hardwick, S.W.; Chirgadze, D.Y.; Worrall, J.A.R.; Blundell, T.L.; Chaplin, A.K.
Using cryo-EM to understand antimycobacterial resistance in the catalase-peroxidase (KatG) from Mycobacterium tuberculosis (2021), Structure, 29, 899-912.e4 .

Search for more literature for 1.11.1.21

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified wild-type and mutant enzymes with bound isoniazid, X-ray diffraction structure determination and analysis at 2.7-3.7 A resolution, dimeric cryo-electronmicroscopic structure of KatGINH. The two protomers are nearly identical, with a root-meansquare deviation (RMSD) value of 0.4 A for Ca atoms when superposing protomer A onto protomer B. There is one notable difference, with protomer B displaying density for residues 206-221. These residues form part of a large loop insertion (LL1), which extends from Glu195 to Asn231 and includes Tyr229 of the MYW catalytic triad. Density for these residues is not observed in protomer A	Mycobacterium tuberculosis

Crystallization (Comment)

Organism

purified wild-type and mutant enzymes with bound isoniazid, X-ray diffraction structure determination and analysis at 2.7-3.7 A resolution, dimeric cryo-electronmicroscopic structure of KatGINH. The two protomers are nearly identical, with a root-meansquare deviation (RMSD) value of 0.4 A for Ca atoms when superposing protomer A onto protomer B. There is one notable difference, with protomer B displaying density for residues 206-221. These residues form part of a large loop insertion (LL1), which extends from Glu195 to Asn231 and includes Tyr229 of the MYW catalytic triad. Density for these residues is not observed in protomer A

Mycobacterium tuberculosis

Protein Variants

Protein Variants	Comment	Organism
T275P	site-directed mutagenesis of the residue from the loop close to the heme binding site. The structure of mutant T275P displays significant areas of disorder compared with the wild-type KatG. Several loops surrounding the heme pocket contain little or no density in either protomer A or B. These disordered regions are identical to those in protomer A of the W107R variant. The loop containing the Thr275 residue (residues 274-329) displays no density and therefore cannot be modeled	Mycobacterium tuberculosis
W107R	site-directed mutagenesis of the catalytic residue, the mutant displays only one heme bound per homodimer of protein. The heme is absent from protomer A and displays significant structural disorder in the vicinity of the heme binding site. Several areas surrounding the heme pocket are difficult to model in protomer A, either displaying minimal or fragmented density. The mutant C-terminal domain of both protomers remains similar to wild-type KatG. The mutant's Arg residue results in disruption of the covalently linked catalytic triad. The loop containing Tyr229, which is part of the MYW catalytic triad is disordered in both protomers, presumably as a consequence of the mutation	Mycobacterium tuberculosis

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Fe2+	the heme group	Mycobacterium tuberculosis

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
161000	-	gel filtration	Mycobacterium tuberculosis

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
2 H2O2	Mycobacterium tuberculosis	-	O2 + 2 H2O	-	?
2 H2O2	Mycobacterium tuberculosis H37Rv	-	O2 + 2 H2O	-	?
2 H2O2	Mycobacterium tuberculosis ATCC 25618	-	O2 + 2 H2O	-	?
isoniazid + H2O2	Mycobacterium tuberculosis	pro-drug activation	?	-	?
isoniazid + H2O2	Mycobacterium tuberculosis H37Rv	pro-drug activation	?	-	?
isoniazid + H2O2	Mycobacterium tuberculosis ATCC 25618	pro-drug activation	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Mycobacterium tuberculosis	P9WIE5	-	-
Mycobacterium tuberculosis ATCC 25618	P9WIE5	-	-
Mycobacterium tuberculosis H37Rv	P9WIE5	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
2 H2O2	-	Mycobacterium tuberculosis	O2 + 2 H2O	-	?
2 H2O2	-	Mycobacterium tuberculosis H37Rv	O2 + 2 H2O	-	?
2 H2O2	-	Mycobacterium tuberculosis ATCC 25618	O2 + 2 H2O	-	?
isoniazid + H2O2	pro-drug activation	Mycobacterium tuberculosis	?	-	?
isoniazid + H2O2	enzyme binding structure analysis, modeling, overview	Mycobacterium tuberculosis	?	-	?
isoniazid + H2O2	pro-drug activation	Mycobacterium tuberculosis H37Rv	?	-	?
isoniazid + H2O2	enzyme binding structure analysis, modeling, overview	Mycobacterium tuberculosis H37Rv	?	-	?
isoniazid + H2O2	pro-drug activation	Mycobacterium tuberculosis ATCC 25618	?	-	?
isoniazid + H2O2	enzyme binding structure analysis, modeling, overview	Mycobacterium tuberculosis ATCC 25618	?	-	?

Subunits

Subunits	Comment	Organism
homodimer	2 * 80000, about sequence calculation	Mycobacterium tuberculosis

Synonyms

Synonyms	Comment	Organism
KatG	-	Mycobacterium tuberculosis
Rv1908c	-	Mycobacterium tuberculosis

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
20	-	assay at	Mycobacterium tuberculosis

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7	-	assay at	Mycobacterium tuberculosis

Cofactor

Cofactor	Comment	Organism	Structure
heme	determination and analysis of the structure of the heme environment of wild-type KatG and KatG bound to isoniazid, overview. The inactivated pro-drug added to the sample is not perturbing the heme site	Mycobacterium tuberculosis

General Information

General Information	Comment	Organism
malfunction	mutations that render the enzyme unable to activate the pro-drug lead to isoniazid (INH) resistance. For two INH resistance variants, W107R and T275P, significant structural disorder relating to heme uptake and retention is the likely cause for INH resistance, dynamics of heme binding are determined by cryo-electronmicroscopy of wild-type and mutant enzymes at 2.7-3.7 A resolution, overview	Mycobacterium tuberculosis
additional information	KatG structure-function analysis, overview	Mycobacterium tuberculosis
physiological function	KatG from Mycobacterium tuberculosis is a catalase-peroxidase that can utilize and degrade hydrogen peroxide (H2O2) either through functioning as a catalase or as a peroxidase. In Mycobacterium tuberculosis, the multifunctional heme enzyme KatG is indispensable for activation of isoniazid (INH), a first-line pro-drug for treatment of tuberculosis. The activated drug species forms an INH-NAD adduct that subsequently triggers anti-tubercular activity	Mycobacterium tuberculosis