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Cloning and characterization of feline islet glucokinase.

Lindbloom-Hawley S, LeCluyse M, Vandersande V, Lushington GH, Schermerhorn T - BMC Vet. Res. (2014)

Bottom Line: The deduced 465 amino acid feline protein has 15 amino acid substitutions not found in other mammalian GK proteins but maintains high structural homology with human GK.Residues crucial for substrate binding and catalysis are completely conserved in the feline protein.Molecular analysis predicts that feline pancreatic GK functions similarly to other mammalian GK proteins.

View Article: PubMed Central - HTML - PubMed

Affiliation: College of Veterinary Medicine, Department of Clinical Sciences, Kansas State University, 1800 Denison Ave, Manhattan, KS 66506-5606, USA. tscherme@vet.ksu.edu.

ABSTRACT

Background: Glucokinase (GK) is a metabolic enzyme encoded by the GCK gene and expressed in glucose-sensitive tissues, principally pancreatic islets cell and hepatocytes. The GK protein acts in pancreatic islets as a "glucose sensor" that couples fluctuations in the blood glucose concentration to changes in cellular function and insulin secretion. GCK and GK have proposed importance in the development and progression of diabetes mellitus and are potential therapeutic targets for diabetes treatment. The study was undertaken to determine the nucleotide sequence of feline pancreatic GK cDNA, predict the amino acid sequence and structure of the feline GK protein, and perform comparative bioinformatic analysis of feline cDNA and protein. Routine PCR techniques were used with cDNA from feline pancreas. Clones were assembled to obtain the full length cDNA. Protein prediction and modeling were performed using bioinformatic tools.

Results: Full-length feline pancreatic GK cDNA contains a 1398 nucleotide coding sequence with high identity to other pancreatic GK cDNAs. The deduced 465 amino acid feline protein has 15 amino acid substitutions not found in other mammalian GK proteins but maintains high structural homology with human GK. Feline pancreatic GK is highly conserved at nucleotide and protein levels. Residues crucial for substrate binding and catalysis are completely conserved in the feline protein.

Conclusion: Molecular analysis predicts that feline pancreatic GK functions similarly to other mammalian GK proteins.

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Related in: MedlinePlus

Alignment of feline GCK with other mammalian GCK sequences. *= Identity at this position in all aligned sequences. : = Conserved substitutions at this position in one or more aligned sequences. . = Semi-conserved substitutions at this position in one or more aligned sequences <space> = non-identity or nonconserved substitution at this position in one or more aligned sequences.
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Figure 2: Alignment of feline GCK with other mammalian GCK sequences. *= Identity at this position in all aligned sequences. : = Conserved substitutions at this position in one or more aligned sequences. . = Semi-conserved substitutions at this position in one or more aligned sequences <space> = non-identity or nonconserved substitution at this position in one or more aligned sequences.

Mentions: The feline pancreatic GK protein deduced from the 1398 bp CDS found in the cDNA is 465 amino acids in length, and has a calculated molecular weight of 52215.67 Daltons or approximately 52 kD (kiloDaltons). Amino acid sequence identities with human, chimpanzee, rat and mouse sequences ranged from 89-94% (Figure 2).


Cloning and characterization of feline islet glucokinase.

Lindbloom-Hawley S, LeCluyse M, Vandersande V, Lushington GH, Schermerhorn T - BMC Vet. Res. (2014)

Alignment of feline GCK with other mammalian GCK sequences. *= Identity at this position in all aligned sequences. : = Conserved substitutions at this position in one or more aligned sequences. . = Semi-conserved substitutions at this position in one or more aligned sequences <space> = non-identity or nonconserved substitution at this position in one or more aligned sequences.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4066705&req=5

Figure 2: Alignment of feline GCK with other mammalian GCK sequences. *= Identity at this position in all aligned sequences. : = Conserved substitutions at this position in one or more aligned sequences. . = Semi-conserved substitutions at this position in one or more aligned sequences <space> = non-identity or nonconserved substitution at this position in one or more aligned sequences.
Mentions: The feline pancreatic GK protein deduced from the 1398 bp CDS found in the cDNA is 465 amino acids in length, and has a calculated molecular weight of 52215.67 Daltons or approximately 52 kD (kiloDaltons). Amino acid sequence identities with human, chimpanzee, rat and mouse sequences ranged from 89-94% (Figure 2).

Bottom Line: The deduced 465 amino acid feline protein has 15 amino acid substitutions not found in other mammalian GK proteins but maintains high structural homology with human GK.Residues crucial for substrate binding and catalysis are completely conserved in the feline protein.Molecular analysis predicts that feline pancreatic GK functions similarly to other mammalian GK proteins.

View Article: PubMed Central - HTML - PubMed

Affiliation: College of Veterinary Medicine, Department of Clinical Sciences, Kansas State University, 1800 Denison Ave, Manhattan, KS 66506-5606, USA. tscherme@vet.ksu.edu.

ABSTRACT

Background: Glucokinase (GK) is a metabolic enzyme encoded by the GCK gene and expressed in glucose-sensitive tissues, principally pancreatic islets cell and hepatocytes. The GK protein acts in pancreatic islets as a "glucose sensor" that couples fluctuations in the blood glucose concentration to changes in cellular function and insulin secretion. GCK and GK have proposed importance in the development and progression of diabetes mellitus and are potential therapeutic targets for diabetes treatment. The study was undertaken to determine the nucleotide sequence of feline pancreatic GK cDNA, predict the amino acid sequence and structure of the feline GK protein, and perform comparative bioinformatic analysis of feline cDNA and protein. Routine PCR techniques were used with cDNA from feline pancreas. Clones were assembled to obtain the full length cDNA. Protein prediction and modeling were performed using bioinformatic tools.

Results: Full-length feline pancreatic GK cDNA contains a 1398 nucleotide coding sequence with high identity to other pancreatic GK cDNAs. The deduced 465 amino acid feline protein has 15 amino acid substitutions not found in other mammalian GK proteins but maintains high structural homology with human GK. Feline pancreatic GK is highly conserved at nucleotide and protein levels. Residues crucial for substrate binding and catalysis are completely conserved in the feline protein.

Conclusion: Molecular analysis predicts that feline pancreatic GK functions similarly to other mammalian GK proteins.

Show MeSH
Related in: MedlinePlus