Literature summary for 3.9.1.2 extracted from

Fuhrmann, J.; Mierzwa, B.; Trentini, D.B.; Spiess, S.; Lehner, A.; Charpentier, E.; Clausen, T.
Structural basis for recognizing phosphoarginine and evolving residue-specific protein phosphatases in gram-positive bacteria (2013), Cell Rep., 3, 1832-1839.

Cloned(Commentary)

Cloned (Comment)	Organism
-	Geobacillus stearothermophilus

Crystallization (Commentary)

Crystallization (Comment)	Organism
in complex with phosphate.The phosphate-binding site is formed by the side chain of Arg13, the backbone amides of the phosphate-loop and the positive end of the macrodipole of helix H1, which together generate a highly positively charged pocket at the bottom of the substrate-binding cleft. The substrate-mimicking Arg149 is sandwiched between Thr11, Asp118, and Phe120 with its guanidinium group hydrogen bonding to Asp118. Structure of mutant C7S in complex with phosphate shows the formation of a covalent phospho-Ser7 adduct indicating that the crystallized mutant mimics the phospho-enzyme intermediate of the dephosphorylation reaction	Geobacillus stearothermophilus

Crystallization (Comment)

Organism

in complex with phosphate.The phosphate-binding site is formed by the side chain of Arg13, the backbone amides of the phosphate-loop and the positive end of the macrodipole of helix H1, which together generate a highly positively charged pocket at the bottom of the substrate-binding cleft. The substrate-mimicking Arg149 is sandwiched between Thr11, Asp118, and Phe120 with its guanidinium group hydrogen bonding to Asp118. Structure of mutant C7S in complex with phosphate shows the formation of a covalent phospho-Ser7 adduct indicating that the crystallized mutant mimics the phospho-enzyme intermediate of the dephosphorylation reaction

Geobacillus stearothermophilus

Protein Variants

Protein Variants	Comment	Organism
C7S	structure in complex with phosphate shows the formation of a covalent phospho-Ser7 adduct indicating that the crystallized mutant mimics the phospho-enzyme intermediate of the dephosphorylation reaction	Geobacillus stearothermophilus
D118A	complete loss of activity	Geobacillus stearothermophilus
F120A	60fold reduction of specific activity	Geobacillus stearothermophilus
T11I	18fold reduction of specific activity with phospho-arginine substrates, marked increasing in the activity toward phospho-tyrosine substrates	Geobacillus stearothermophilus
T11V	18fold reduction of specific activity with phospho-arginine substrates, marked increasing in the activity toward phospho-tyrosine substrates	Geobacillus stearothermophilus

Organism

Organism	UniProt	Comment	Textmining
Geobacillus stearothermophilus	S0F332	-	-

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
KpRGGGGYIKIIKV + H2O	hydrolysis of the phosphoramidate bond of phospho-arginine residues in peptides	Geobacillus stearothermophilus	KRGGGGYIKIIKV + phosphate	-	?