Comprehensive analysis of CXXX sequence space reveals that S. cerevisiae GGTase-I mainly relies on a 2 X substrate determinants.

Autor: Sarkar A; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602., Hildebrandt ER; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602., Patel KV; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602., Mai ET; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602., Shah SS; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602., Kim JH; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602., Schmidt WK; Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, 30602.
Jazyk: angličtina
Zdroj: BioRxiv : the preprint server for biology [bioRxiv] 2024 Mar 04. Date of Electronic Publication: 2024 Mar 04.
DOI: 10.1101/2024.03.04.583369
Abstrakt: Many proteins undergo a post-translational lipid attachment, which increases their hydrophobicity, thus strengthening their membrane association properties or aiding in protein interactions. Geranylgeranyltransferase-I (GGTase-I) is an enzyme involved in a three-step post-translational modification (PTM) pathway that attaches a 20-carbon lipid group called geranylgeranyl at the carboxy-terminal cysteine of proteins ending in a canonical CaaL motif (C - cysteine, a - aliphatic, L - often leucine, but can be phenylalanine, isoleucine, methionine, or valine). Genetic approaches involving two distinct reporters were employed in this study to assess S. cerevisiae GGTase-I specificity, for which limited data exists, towards all 8000 CXXX combinations. Orthogonal biochemical analyses and structure-based alignments were also performed to better understand the features required for optimal target interaction. These approaches indicate that yeast GGTase-I best modifies the Cxa[L/F/I/M/V] sequence that resembles but is not an exact match for the canonical CaaL motif. We also observed that minor modification of non-canonical sequences is possible. A consistent feature associated with well-modified sequences was the presence of a non-polar a 2 residue and a hydrophobic terminal residue, which are features recognized by mammalian GGTase-I. These results thus support that mammalian and yeast GGTase-I exhibit considerable shared specificity.
Competing Interests: Conflict of interest The authors have declared no competing interests exist.
Databáze: MEDLINE
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