PUBLICATION
Structure Dissection of Zebrafish Progranulins Identifies a Well-Folded Granulin/Epithelin Module Protein with pro-Cell Survival Activities
- Authors
- Wang, P., Chitramuthu, B., Bateman, A., Bennett, H.P.J., Xu, P., Ni, F.
- ID
- ZDB-PUB-180509-14
- Date
- 2018
- Source
- Protein science : a publication of the Protein Society 27(8): 1476-1490 (Journal)
- Registered Authors
- Keywords
- 3D structure, NMR spectroscopy, granulin/epithelin module, hairpin stack fold, progranulin, zebrafish
- MeSH Terms
-
- Animals
- Cell Line
- Cell Survival/drug effects*
- Fish Proteins/chemistry*
- Fish Proteins/genetics
- Fish Proteins/metabolism
- Fish Proteins/pharmacology*
- Mice
- Neuroprotective Agents/chemistry
- Neuroprotective Agents/pharmacology
- Progranulins/chemistry*
- Progranulins/genetics
- Progranulins/metabolism
- Progranulins/pharmacology*
- Protein Folding
- Recombinant Fusion Proteins/chemistry
- Recombinant Fusion Proteins/genetics
- Recombinant Fusion Proteins/metabolism
- Recombinant Fusion Proteins/pharmacology
- Zebrafish
- PubMed
- 29732682 Full text @ Protein Sci.
Citation
Wang, P., Chitramuthu, B., Bateman, A., Bennett, H.P.J., Xu, P., Ni, F. (2018) Structure Dissection of Zebrafish Progranulins Identifies a Well-Folded Granulin/Epithelin Module Protein with pro-Cell Survival Activities. Protein science : a publication of the Protein Society. 27(8):1476-1490.
Abstract
The ancient and pluripotent progranulins contain multiple repeats of a cysteine-rich sequence motif of ∼60 amino acids, called the granulin/epithelin module (GEM) with a prototypic structure of four β-hairpins zipped together by six inter-hairpin disulfide bonds. Prevalence of this disulfide-enforced structure is assessed here by an expression screening of 19 unique GEM sequences of the four progranulins in the zebrafish genome, progranulins 1, 2, A and B. While a majority of the expressed GEM peptides did not exhibit uniquely folded conformations, module AaE from progranulin A and AbB from progranulin B were found to fold into the protopypic 4-hairpin structure along with disulfide formation. Module AaE has the most-rigid three-dimensional structure with all four β-hairpins defined using high-resolution (H-15 N) NMR spectroscopy, including 492 inter-proton nuclear Overhauser effects, 23 3 J(HN,Hα ) coupling constants, 22 hydrogen bonds as well as 45 residual dipolar coupling constants. Three-dimensional structure of AaE and the partially folded AbB re-iterate the conformational stability of the N-terminal stack of two beta-hairpins and varying degrees of structural flexibility for the C-terminal half of the 4-hairpin global fold of the GEM repeat. A cell-based assay demonstrated a functional activity for the zebrafish granulin AaE in promoting the survival of neuronal cells, similarly to what has been found for the corresponding granulin E module in human progranulin. Finally, this work highlights the remaining challenges in structure-activity studies of proteins containing the GEM repeats, due to the apparent prevalence of structural disorder in GEM motifs despite potentially a high density of intramolecular disulfide bonds.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping