Cannabinoid receptor interacting protein 1a interacts with myristoylated Gαi N terminus via a unique gapped β-barrel structure

• Published in: The Journal of biological chemistry Vol. 297; no. 3; p. 101099
• Main Authors: Booth, William T., Clodfelter, Jill E., Leone-Kabler, Sandra, Hughes, Erin K., Eldeeb, Khalil, Howlett, Allyn C., Lowther, W. Todd
• Format: Journal Article
• Language: English
• Published: Elsevier Inc 01-09-2021; American Society for Biochemistry and Molecular Biology
• Subjects: Accelerated Communication; endocannabinoid system; fatty acid–binding protein; G protein–coupled receptors; guanine nucleotide dissociation inhibitor; small G proteins; guanine nucleotide dissociation inhibitor; fatty acid–binding protein; PDE6δ; FABP; endocannabinoid system; G protein–coupled receptors; BRIL; CRIP1a; GDI; T4L; small G proteins; CB1R
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/j.jbc.2021.101099

Cannabinoid receptor interacting protein 1a (CRIP1a) modulates CB1 cannabinoid receptor G-protein coupling in part by altering the selectivity for Gαi subtype activation, but the molecular basis for this function of CRIP1a is not known. We report herein the first structure of CRIP1a at a resolution of 1.55 Å. CRIP1a exhibits a 10-stranded and antiparallel β-barrel with an interior comprised of conserved hydrophobic residues and loops at the bottom and a short helical cap at the top to exclude solvent. The β-barrel has a gap between strands β8 and β10, which deviates from β-sandwich fatty acid–binding proteins that carry endocannabinoid compounds and the Rho-guanine nucleotide dissociation inhibitor predicted by computational threading algorithms. The structural homology search program DALI identified CRIP1a as homologous to a family of lipidated-protein carriers that includes phosphodiesterase 6 delta subunit and Unc119. Comparison with these proteins suggests that CRIP1a may carry two possible types of cargo: either (i) like phosphodiesterase 6 delta subunit, cargo with a farnesyl moiety that enters from the top of the β-barrel to occupy the hydrophobic interior or (ii) like Unc119, cargo with a palmitoyl or a myristoyl moiety that enters from the side where the missing β-strand creates an opening to the hydrophobic pocket. Fluorescence polarization analysis demonstrated CRIP1a binding of an N-terminally myristoylated 9-mer peptide mimicking the Gαi N terminus. However, CRIP1a could not bind the nonmyristolyated Gαi peptide or cargo of homologs. Thus, binding of CRIP1a to Gαi proteins represents a novel mechanism to regulate cell signaling initiated by the CB1 receptor.