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Literature summary for 6.5.1.3 extracted from

  • Unciuleac, M.C.; Shuman, S.
    Characterization of a novel eukaryal nick-sealing RNA ligase from Naegleria gruberi (2015), RNA, 21, 824-832 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 6.5.1.3

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21(DE3) cells Naegleria gruberi

Protein Variants

Protein Variants Comment Organism
K170A inactive Naegleria gruberi
K170H inactive Naegleria gruberi
K170M inactive Naegleria gruberi
K170Q inactive Naegleria gruberi

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+ enzyme activity is optimal at 5 mM Mg2+ Naegleria gruberi
Mn2+ enzyme activity is optimal at 2.5 mM Mn2+ Naegleria gruberi
additional information calcium, cobalt, copper, cadmium, nickel, and zinc are ineffective Naegleria gruberi

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
38000
-
 gel filtration Naegleria gruberi

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + (ribonucleotide)n-3'-hydroxyl + 5'-phospho-(ribonucleotide)m Naegleria gruberi
-
 (ribonucleotide)n+m + AMP + diphosphate
-
 ?
ATP + [RNA ligase]-L-lysine Naegleria gruberi
-
 [RNA ligase]-N6-(5'-adenylyl)-L-lysine + diphosphate
-
 ?

Organism

Organism UniProt Comment Textmining
Naegleria gruberi
-
-
-

Purification (Commentary)

Purification (Comment) Organism
Ni-NTA agarose column chromatography, and DEAE Sephacel gel filtration Naegleria gruberi

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + (ribonucleotide)n-3'-hydroxyl + 5'-phospho-(ribonucleotide)m
-
 Naegleria gruberi (ribonucleotide)n+m + AMP + diphosphate
-
 ?
ATP + [RNA ligase]-L-lysine
-
 Naegleria gruberi [RNA ligase]-N6-(5'-adenylyl)-L-lysine + diphosphate
-
 ?
additional information the enzyme is unreactive with a 5'-32P-labeled single-strand 18-mer pDNA substrate Naegleria gruberi ?
-
 ?
additional information the enzyme seals nicked 3'-OH/5'-PO4 duplexes in which the 3'-OH strand is RNA. The enzyme entails reaction with ATP to form a covalent enzyme-AMP intermediate, transfer of AMP to the nick 5'-PO4, and attack of the RNA 3'-OH on the adenylylated nick to form a 3'-5' phosphodiester Naegleria gruberi ?
-
 ?

Subunits

Subunits Comment Organism
monomer 1 * 38000, SDS-PAGE Naegleria gruberi

Synonyms

Synonyms Comment Organism
ATP-dependent RNA ligase
-
 Naegleria gruberi
Rnl5
-
 Naegleria gruberi

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
6
-
-
 Naegleria gruberi

pH Range

pH Minimum pH Maximum Comment Organism
5.5 8.5 nick sealing activity is optimal at pH 5.5 to 6.5 and declined sharply at pH 8.5 or above Naegleria gruberi