Skip to main content

Biochemical characterization of purified mammalian ARL13B protein indicates that it is an atypical GTPase and ARL3 guanine nucleotide exchange factor (GEF).

Publication ,  Journal Article
Ivanova, AA; Caspary, T; Seyfried, NT; Duong, DM; West, AB; Liu, Z; Kahn, RA
Published in: J Biol Chem
June 30, 2017

Primary cilia play central roles in signaling during metazoan development. Several key regulators of ciliogenesis and ciliary signaling are mutated in humans, resulting in a number of ciliopathies, including Joubert syndrome (JS). ARL13B is a ciliary GTPase with at least three missense mutations identified in JS patients. ARL13B is a member of the ADP ribosylation factor family of regulatory GTPases, but is atypical in having a non-homologous, C-terminal domain of ∼20 kDa and at least one key residue difference in the consensus GTP-binding motifs. For these reasons, and to establish a solid biochemical basis on which to begin to model its actions in cells and animals, we developed preparations of purified, recombinant, murine Arl13b protein. We report results from assays for solution-based nucleotide binding, intrinsic and GTPase-activating protein-stimulated GTPase, and ARL3 guanine nucleotide exchange factor activities. Biochemical analyses of three human missense mutations found in JS and of two consensus GTPase motifs reinforce the atypical properties of this regulatory GTPase. We also discovered that murine Arl13b is a substrate for casein kinase 2, a contaminant in our preparation from human embryonic kidney cells. This activity, and the ability of casein kinase 2 to use GTP as a phosphate donor, may be a source of differences between our data and previously published results. These results provide a solid framework for further research into ARL13B on which to develop models for the actions of this clinically important cell regulator.

Duke Scholars

Altmetric Attention Stats
Dimensions Citation Stats

Published In

J Biol Chem

DOI

EISSN

1083-351X

Publication Date

June 30, 2017

Volume

292

Issue

26

Start / End Page

11091 / 11108

Location

United States

Related Subject Headings

  • Retina
  • Mutation, Missense
  • Mice
  • Kidney Diseases, Cystic
  • Humans
  • Eye Abnormalities
  • Cerebellum
  • Casein Kinase II
  • Biochemistry & Molecular Biology
  • Animals
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Ivanova, A. A., Caspary, T., Seyfried, N. T., Duong, D. M., West, A. B., Liu, Z., & Kahn, R. A. (2017). Biochemical characterization of purified mammalian ARL13B protein indicates that it is an atypical GTPase and ARL3 guanine nucleotide exchange factor (GEF). J Biol Chem, 292(26), 11091–11108. https://doi.org/10.1074/jbc.M117.784025
Ivanova, Anna A., Tamara Caspary, Nicholas T. Seyfried, Duc M. Duong, Andrew B. West, Zhiyong Liu, and Richard A. Kahn. “Biochemical characterization of purified mammalian ARL13B protein indicates that it is an atypical GTPase and ARL3 guanine nucleotide exchange factor (GEF).J Biol Chem 292, no. 26 (June 30, 2017): 11091–108. https://doi.org/10.1074/jbc.M117.784025.
Ivanova AA, Caspary T, Seyfried NT, Duong DM, West AB, Liu Z, et al. Biochemical characterization of purified mammalian ARL13B protein indicates that it is an atypical GTPase and ARL3 guanine nucleotide exchange factor (GEF). J Biol Chem. 2017 Jun 30;292(26):11091–108.
Ivanova, Anna A., et al. “Biochemical characterization of purified mammalian ARL13B protein indicates that it is an atypical GTPase and ARL3 guanine nucleotide exchange factor (GEF).J Biol Chem, vol. 292, no. 26, June 2017, pp. 11091–108. Pubmed, doi:10.1074/jbc.M117.784025.
Ivanova AA, Caspary T, Seyfried NT, Duong DM, West AB, Liu Z, Kahn RA. Biochemical characterization of purified mammalian ARL13B protein indicates that it is an atypical GTPase and ARL3 guanine nucleotide exchange factor (GEF). J Biol Chem. 2017 Jun 30;292(26):11091–11108.

Published In

J Biol Chem

DOI

EISSN

1083-351X

Publication Date

June 30, 2017

Volume

292

Issue

26

Start / End Page

11091 / 11108

Location

United States

Related Subject Headings

  • Retina
  • Mutation, Missense
  • Mice
  • Kidney Diseases, Cystic
  • Humans
  • Eye Abnormalities
  • Cerebellum
  • Casein Kinase II
  • Biochemistry & Molecular Biology
  • Animals