The Crystal Structure of Choline Kinase Reveals a Eukaryotic Protein Kinase Fold

Peisach, Daniel; Gee, Patricia; Kent, Claudia; Xu, Zhaohui

doi:10.1016/s0969-2126(03)00094-7

Cited by 69 publications

(78 citation statements)

References 49 publications

(2 reference statements)

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“…NPL structures do have in common their high mobility, and their conformations undergo rearrangement upon binding of ATP. The loop structure of CKA-2 is somewhat similar to that of APH(3Ј)-IIIa in terms of both primary and tertiary structure (22,38). First, they both have a GMS sequence instead of the classical GXGXXG sequence in most ePKs (Fig.…”

Section: Fig 3 the Mgmentioning

confidence: 85%

“…The crystal structure of CKA-2 demonstrates that Trp-387 is located in a pocket that is not found in the active sites of ePKs and APH(3Ј)-IIIa (Fig. 6) (22). Trp-387 lies close to the binding site of the ␥-phosphate of ATP; two other aromatic residues, Trp-390 and Tyr-304, are close to the Trp-387 in this pocket structure.…”

Section: Fig 3 the Mgmentioning

confidence: 97%

“…The crystal structure shows that four of these residues are involved in mutual interactions. The side chain of Asp-313 hydrogenbonds with the main chain of His-253 and Asn-254, whereas His-181 interacts with the carboxylate group of Asp-313 (22). This evidence suggests that these residues have structural roles in maintaining the proper conformation of the enzyme.…”

Section: Fig 3 the Mgmentioning

confidence: 98%

“…This enzyme is as catalytically active as choline kinase from mammals and yeast and is quite similar to them in primary structure. Moreover, CKA-2 has been amenable to crystallization, and the crystal structure of CKA-2 has been determined recently (22). Surprisingly, the three-dimensional structure of CKA-2 is quite similar to those of protein kinases and aminoglycoside phosphotransferases, despite very low similarity in amino acid sequences among these enzymes.…”

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confidence: 99%

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Identification of Critical Residues of Choline Kinase A2 from Caenorhabditis elegans

Yuan

Kent

2004

Journal of Biological Chemistry

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Choline kinase catalyzes the phosphorylation of choline by ATP, the first committed step in the CDP-choline pathway for phosphatidylcholine biosynthesis. To begin to elucidate the mechanism of catalysis by this enzyme, choline kinase A-2 from Caenorhabditis elegans was analyzed by systematic mutagenesis of highly conserved residues followed by analysis of kinetic and structural parameters. Specifically, mutants were analyzed with respect to K m and k cat values for each substrate and Mg 2؉ , inhibitory constants for Mg 2؉ and Ca 2؉ , secondary structure as monitored by circular dichroism, and sensitivity to unfolding in guanidinium hydrochloride. The most severe impairment of catalysis occurred with the modification of Asp-255 and Asn-260, which are located in the conserved Brenner's phosphotransferase motif, and Asp-301 and Glu-303, in the signature choline kinase motif. For example, mutation of Asp-255 or Asp-301 to Ala eliminated detectable catalytic activity, and mutation of Asn-260 and Glu-303 to Ala decreased k cat by 300-and 10-fold, respectively. Additionally, the K m for Mg 2؉ for mutants N260A and E303A was approximately 30-fold higher than that of wild type. Several other residues (Ser-86, Arg-111, Glu-125, and Trp-387) were identified as being important: Catalytic efficiencies (k cat /K m ) for the enzymes in which these residues were mutated to Ala were reduced to 2-25% of wild type. The high degree of structural similarity among choline kinase A-2, aminoglycoside phosphotransferases, and protein kinases, together with the results from this mutational analysis, indicates it is likely that these conserved residues are located at the catalytic core of choline kinase.

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Section: Fig 3 the Mgmentioning

confidence: 85%

Section: Fig 3 the Mgmentioning

confidence: 97%

Section: Fig 3 the Mgmentioning

confidence: 98%

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confidence: 99%

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Identification of Critical Residues of Choline Kinase A2 from Caenorhabditis elegans

Yuan

Kent

2004

Journal of Biological Chemistry

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“…Entries were not provided when no significant similarities were detected and no BLAST score could be calculated by the BLAST algorithm. tein kinases (16). As a group of proteins, tRNA synthetases appeared among the top hits in all six species employed for BLAST search.…”

Section: Alignment Of Human Proteinmentioning

confidence: 99%

Evolutionary Ancestry of Eukaryotic Protein Kinases and Choline Kinases

Lai

Safaei

Pelech

2016

Journal of Biological Chemistry

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The reversible phosphorylation of proteins catalyzed by protein kinases in eukaryotes supports an important role for eukaryotic protein kinases (ePKs) in the emergence of nucleated cells in the third superkingdom of life. Choline kinases (ChKs) could also be critical in the early evolution of eukaryotes, because of their function in the biosynthesis of phosphatidylcholine, which is unique to eukaryotic membranes. However, the genomic origins of ePKs and ChKs are unclear. The high degeneracy of protein sequences and broad expansion of ePK families have made this fundamental question difficult to answer. In this study, we identified two class-I aminoacyl-tRNA synthetases with high similarities to consensus amino acid sequences of human protein-serine/threonine kinases. Comparisons of primary and tertiary structures supported that ePKs and ChKs evolved from a common ancestor related to glutaminyl aminoacyl-tRNA synthetases, which may have been one of the key factors in the successful of emergence of ancient eukaryotic cells from bacterial colonies.Protein kinases play a pivotal role in communicating intracellular signals in eukaryotes. The family of eukaryotic protein kinases (ePKs) 3 comprises at least 568 human members, which accounts for more than 2% of the protein coding genes of the entire human genome (1). These kinases are highly conserved both in their primary amino acid sequences (2) and in the threedimensional structures (3) of their catalytic domains. Because of the central regulatory roles and the high conservation of the ePKs, the ancestry of these enzymes has become an important question in the study of the evolution of eukaryotic organisms.The majority of the kinases among the ePKs are responsible for the phosphorylation of proteins on serine or threonine residues, whereas a smaller group of protein kinases catalyzes their tyrosine phosphorylation. This branch of protein-tyrosine kinases (PTKs) arose from protein-serine/threonine kinases (STKs), which is believed to be an important event in early metazoan evolution (4, 5). Of all the STKs, there is another lumped group of diverse kinases that are described as atypical protein kinases. With little sequence identity and structural similarity to typical protein kinases, these atypical protein kinases are suggested to have diverged early in evolution and have distinct evolutionary histories (6, 7). Despite the atypical protein kinases and recently derived PTKs, the rest of the typical protein kinases constitutes a major lineage in protein kinase evolution.Eukaryotic life is believed to have evolved between 1.7 and 2.7 billion years ago, and no living representatives of the earliest eukaryotes survive today. Consequently, the actual origin of protein kinases is difficult to establish with a high degree of confidence. Firstly, protein sequences are highly degenerate, which makes the detection of sequence similarities difficult even at the superfamily level (8). Secondly, the ePKs comprise a group of very broadly expanded proteins. Loss and expansion of kin...

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From yeast to humans – roles of the Kennedy pathway for phosphatidylcholine synthesis

McMaster

2017

FEBS Letters

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The major phospholipid present in most eukaryotic membranes is phosphatidylcholine (PC), comprising~50% of phospholipid content. PC metabolic pathways are highly conserved from yeast to humans. The main pathway for the synthesis of PC is the Kennedy (CDP-choline) pathway. In this pathway, choline is converted to phosphocholine by choline kinase, phosphocholine is metabolized to CDP-choline by the rate-determining enzyme for this pathway, CTP:phosphocholine cytidylyltransferase, and cholinephosphotransferase condenses CDP-choline with diacylglycerol to produce PC. This Review discusses how PC synthesis via the Kennedy pathway is regulated, its role in cellular and biological processes, as well as diseases known to be associated with defects in PC synthesis. Finally, we present the first model for the making of a membrane via PC synthesis.

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The Crystal Structure of Choline Kinase Reveals a Eukaryotic Protein Kinase Fold

Cited by 69 publications

References 49 publications

Identification of Critical Residues of Choline Kinase A2 from Caenorhabditis elegans

Identification of Critical Residues of Choline Kinase A2 from Caenorhabditis elegans

Evolutionary Ancestry of Eukaryotic Protein Kinases and Choline Kinases

From yeast to humans – roles of the Kennedy pathway for phosphatidylcholine synthesis

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