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Literature summary extracted from

  • Newmister, S.; Otte, M.; Escalante-Semerena, J.; Rayment, I.
    Structure and Mutational Analysis of the Archaeal GTP:AdoCbi-P Guanylyltransferase (CobY) from Methanocaldococcus jannaschii: Insights into GTP Binding and Dimerization (2011), Biochemistry, 50, 5301-5313.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.7.7.62 expressed in Escherichia coli Methanocaldococcus jannaschii

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.7.7.62 three-dimensional crystal structure of the guanylyltransferase (CobY) enzyme (mutant G153D) from Methanocaldococcus jannaschii (MjCobY)1 in complex with GTP at 2.8 A resolution is reported Methanocaldococcus jannaschii

Protein Variants

EC Number Protein Variants Comment Organism
2.7.7.62 D101G in vitro activity relative to wild-type: 71%, monomer/dimer ratio: 34/66 Methanocaldococcus jannaschii
2.7.7.62 D83G in vitro activity relative to wild-type: 100%, monomer/dimer ratio: 82/18 Methanocaldococcus jannaschii
2.7.7.62 E18D in vitro activity relative to wild-type: 56%, monomer/dimer ratio: 40/60 Methanocaldococcus jannaschii
2.7.7.62 G153D crystal structure of mutant is determined at 2.8 A: The protein crystallizes in the hexagonal space group P31 with 4 chains in the asymmetric unit. Mutant shows reduced activity and is composed of a monomer. Results from isothermal titration calorimetry experiments show that MjCobYG153D has 10fold higher affinity for GTP than wild-type, but fails to bind the corrinoid substrate Methanocaldococcus jannaschii
2.7.7.62 G153D in vitro activity relative to wild-type: 2%, monomer/dimer ratio: 54/46 Methanocaldococcus jannaschii
2.7.7.62 G8D mutant non-functional, monomer/dimer ratio: 33/67 Methanocaldococcus jannaschii
2.7.7.62 I144D/F146D/Y163R monomer/dimer/aggregate ratio: 77/23/0. Specific activity (pmol/min/microgram): 156 (monomer), 49 (dimer) Methanocaldococcus jannaschii
2.7.7.62 K19A in vitro activity relative to wild-type: 16%, monomer/dimer ratio: 36/64 Methanocaldococcus jannaschii
2.7.7.62 L38D/K41D monomer/dimer/aggregate ratio: 53/25/22. Specific activity (pmol/min/microgram): 125 (monomer), 120 (dimer), 78 (aggregate) Methanocaldococcus jannaschii
2.7.7.62 L38K/K41D monomer/dimer/aggregate ratio: 52/26/22. Specific activity (pmol/min/microgram): 124 (monomer), 119 (dimer), 85 (aggregate) Methanocaldococcus jannaschii
2.7.7.62 N177R in vitro activity relative to wild-type: 5%, monomer/dimer ratio: not determined Methanocaldococcus jannaschii
2.7.7.62 N179D in vitro activity relative to wild-type: 95%, monomer/dimer ratio: not determined Methanocaldococcus jannaschii
2.7.7.62 P20A in vitro activity relative to wild-type: 93%, monomer/dimer ratio: 63/37 Methanocaldococcus jannaschii
2.7.7.62 P54A in vitro activity relative to wild-type: 62%, monomer/dimer ratio: 50/50 Methanocaldococcus jannaschii
2.7.7.62 P94A in vitro activity relative to wild-type: 43%, monomer/dimer ratio: 52/48 Methanocaldococcus jannaschii
2.7.7.62 R13A in vitro activity relative to wild-type: 15%, monomer/dimer ratio: 14/86 Methanocaldococcus jannaschii
2.7.7.62 S100A in vitro activity relative to wild-type: 100%, monomer/dimer ratio: not determined Methanocaldococcus jannaschii
2.7.7.62 S50A in vitro activity relative to wild-type: 100%, monomer/dimer ratio: not determined Methanocaldococcus jannaschii
2.7.7.62 T180A in vitro activity relative to wild-type: 73%, monomer/dimer ratio: not determined Methanocaldococcus jannaschii
2.7.7.62 T53S in vitro activity relative to wild-type: 84%, monomer/dimer ratio: 76/24 Methanocaldococcus jannaschii
2.7.7.62 T56A in vitro activity relative to wild-type: 102%, monomer/dimer ratio: not determined Methanocaldococcus jannaschii
2.7.7.62 Y80F in vitro activity relative to wild-type: 65%, monomer/dimer ratio: 74/26 Methanocaldococcus jannaschii

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.7.7.62 GTP + adenosylcobinamide phosphate Methanocaldococcus jannaschii
-
diphosphate + adenosylcobinamide-GDP
-
?
2.7.7.62 GTP + adenosylcobinamide phosphate Methanocaldococcus jannaschii DSM 2661
-
diphosphate + adenosylcobinamide-GDP
-
?

Organism

EC Number Organism UniProt Comment Textmining
2.7.7.62 Methanocaldococcus jannaschii Q58517
-
-
2.7.7.62 Methanocaldococcus jannaschii DSM 2661 Q58517
-
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.7.7.62 GTP + adenosylcobinamide phosphate
-
Methanocaldococcus jannaschii diphosphate + adenosylcobinamide-GDP
-
?
2.7.7.62 GTP + adenosylcobinamide phosphate
-
Methanocaldococcus jannaschii DSM 2661 diphosphate + adenosylcobinamide-GDP
-
?

Subunits

EC Number Subunits Comment Organism
2.7.7.62 dimer MjCobYWT is comprised of a population of dimers and monomers. CobY is active in both the monomeric and dimeric forms Methanocaldococcus jannaschii
2.7.7.62 monomer MjCobYWT is comprised of a population of dimers and monomers. CobY is active in both the monomeric and dimeric forms Methanocaldococcus jannaschii

Synonyms

EC Number Synonyms Comment Organism
2.7.7.62 CobY
-
Methanocaldococcus jannaschii
2.7.7.62 MjCobY
-
Methanocaldococcus jannaschii

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.7.7.62 37
-
assay at Methanocaldococcus jannaschii

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.7.7.62 7.9
-
assay at Methanocaldococcus jannaschii