PUBLICATION
Expression, purification and oligomerization of the S-adenosylmethionine transporter
- Authors
- Wang, D., Liu, M., Li, X., Wang, X., Shen, Y.
- ID
- ZDB-PUB-210123-32
- Date
- 2020
- Source
- Protein Expression and Purification 173: 105648 (Journal)
- Registered Authors
- Keywords
- Expression, Membrane protein, Purification, S-Adenosylmethionine, SAMC
- MeSH Terms
-
- Amino Acid Transport Systems*/biosynthesis
- Amino Acid Transport Systems*/chemistry
- Amino Acid Transport Systems*/genetics
- Amino Acid Transport Systems*/isolation & purification
- Animals
- Gene Expression*
- Humans
- Male
- Protein Multimerization*
- Recombinant Proteins/biosynthesis
- Recombinant Proteins/chemistry
- Recombinant Proteins/genetics
- Recombinant Proteins/isolation & purification
- Zebrafish/genetics*
- Zebrafish/metabolism
- Zebrafish Proteins*/biosynthesis
- Zebrafish Proteins*/chemistry
- Zebrafish Proteins*/genetics
- Zebrafish Proteins*/isolation & purification
- PubMed
- 32335303 Full text @ Protein Expr. Purif.
Citation
Wang, D., Liu, M., Li, X., Wang, X., Shen, Y. (2020) Expression, purification and oligomerization of the S-adenosylmethionine transporter. Protein Expression and Purification. 173:105648.
Abstract
The S-adenosylmethionine carrier (SAMC) is a membrane transport protein located on the inner membrane of mitochondria that catalyzes the import of S-adenosylmethionine (SAM) into the mitochondrial matrix. SAMC mutations can cause a series of mitochondrial defects, including those affecting RNA stability, protein modification, mitochondrial translation and biosynthesis. Here, we describe the expression, purification and oligomerization of SAMC. The SAMC genes from three species were cloned into a eukaryotic expression vector with a GFP tag, and confocal microscopy analysis showed that these SAMCs were localized to mitochondria. A BacMam expression system was used for the expression of D. rerio SAMC with a FLAG tag. A size-exclusion chromatography analysis showed that SAMC may form a hexamer. A negative-staining electron microscopy analysis showed that SAMC formed tiny uniform particles and also confirmed the oligomerization of SAMC.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping