What question did this study set out to answer?

To investigate the physical interaction, macromolecular organization, and intracellular localization of the brefeldin A-inhibited guanine nucleotide-exchange proteins BIG1 and BIG2.

March 14, 2000Proceedings of the National Academy of Sciences

Identification and localization of two brefeldin A-inhibited guanine nucleotide-exchange proteins for ADP-ribosylation factors in a macromolecular complex

View Full Paper

Ask AI

Bookmark

Authors

RYRyoichi YamajiMetropolitan University RARonald AdamikNational Institutes of Health KTKazuyo TakedaCenter for Biologics Evaluation and Research

Discussion

Member takes

Implication

Laboratory study reveals BIG1 and BIG2 form a macromolecular complex associated with Golgi membranes in mammalian cells, suggesting coordinated regulation of membrane trafficking.

Key Points

To investigate the physical interaction, macromolecular organization, and intracellular localization of the brefeldin A-inhibited guanine nucleotide-exchange proteins BIG1 and BIG2.
Purified protein complexes from bovine brain cytosol and analyzed HeLa S3 cell lysates using gel filtration chromatography.
Performed subcellular fractionation and co-immunoprecipitation from cytosolic and microsomal fractions using specific anti-peptide antibodies against BIG1 and BIG2.
Visualized endogenous and coexpressed proteins in HeLa S3 and HepG2 cells via immunofluorescence microscopy alongside Golgi markers.
BIG1 (200 kDa) and BIG2 (190 kDa) copurified in macromolecular complexes with molecular masses exceeding 670 kDa and cofractionated with the Golgi marker beta-COP.
Co-immunoprecipitation showed approximately 70% of BIG2 coprecipitated with 100% of BIG1, and approximately 75% of BIG1 coprecipitated with 100% of BIG2.
Both proteins showed a punctate cytosolic distribution and concentrated perinuclearly, partially colocalizing with Golgi-specific 58K protein and gamma-adaptin.

Cite This Study

Yamaji et al. (2000) studied this question.

synapsesocial.com/papers/6a85cc5d0b32cc5dc4cb35db https://doi.org/10.1073/pnas.97.6.2567

View Full Paper

Ask AI

Bookmark

Implication

Laboratory study reveals BIG1 and BIG2 form a macromolecular complex associated with Golgi membranes in mammalian cells, suggesting coordinated regulation of membrane trafficking.

Identification and localization of two brefeldin A-inhibited guanine nucleotide-exchange proteins for ADP-ribosylation factors in a macromolecular complex

Discussion

Member takes

Implication

Key Points

Cite This Study

Identification and localization of two brefeldin A-inhibited guanine nucleotide-exchange proteins for ADP-ribosylation factors in a macromolecular complex

Discussion

Member takes

Implication

Key Points

Cite This Study

Also Consider

Also Consider