The N and C Termini of ZO-1 Are Surrounded by Distinct Proteins and Functional Protein Networks

Itallie, Christina M. Van; Aponte, Angel; Tietgens, Amber Jean; Guček, Marjan; Fredriksson, Karin; Anderson, James M.

doi:10.1074/jbc.m113.466193

Cited by 108 publications

(128 citation statements)

References 95 publications

(60 reference statements)

Supporting

Mentioning

118

Contrasting

Order By: Relevance

“…Similar to what we had found using this method with ZO-1 (Van Itallie et al, 2013), our results suggest that E-cadherinbiotin-ligase (EcadBL) identifies a large number of known functionally relevant proteins as well as proteins previously not known to be near E-cadherin and which might provide novel insights about E-cadherin function. The most abundant proteins identified were catenins, including a-E-catenin, b-catenin, p120 catenin and plakoglobin, and unexpectedly, a-N-catenin and a-Tcatenin.…”

Section: Introductionsupporting

confidence: 72%

“…2A); a prominent band at ,135 kDa was detected with an anti-E-cadherin antibody and is likely the EcadBL fusion protein. Unlike the results obtained with ZO-1 biotin ligase fusions (Van Itallie et al, 2013), the E-cadherin fusion protein was not the most abundant biotinylated protein in these cells, possibly because the E-cadherin intracellular domain available for self-biotinylation contains only five lysines. Three similar Coomassie-stained protein gel lanes from independent labeling experiments and streptavidin purifications were each cut into 12 bands (top to bottom) and used for proteomic analysis by mass spectrometry.…”

Section: Ecadbl Fusion Protein Localizes To Cell-cell Contacts In MDCcontrasting

confidence: 46%

“…LPP, a LIM-domain-containing member of zyxin family, is identified as an abundant proximal protein One protein, lipoma preferred partner (LPP, rank 30) was of particular interest because it was also among the more abundant proteins tagged by the biotin ligase ZO-1 fusion protein (rank 36; Van Itallie et al, 2013). E-cadherin is essential not only in adherens junctions, but is also required for normal tight junction formation (Capaldo and Macara, 2007).…”

Section: Ecadbl Fusion Protein Localizes To Cell-cell Contacts In MDCmentioning

confidence: 99%

See 2 more Smart Citations

Biotin ligase tagging identifies proteins proximal to E-cadherin, including lipoma preferred partner, a regulator of epithelial cell-cell and cell-substrate adhesion

Itallie

Tietgens

Aponte

et al. 2013

Journal of Cell Science

Self Cite

116

View full text Add to dashboard Cite

Known proteins associated with the cell-adhesion protein Ecadherin include catenins and proteins involved in signaling, trafficking and actin organization. However, the list of identified adherens junction proteins is likely to be incomplete, limiting investigation into this essential cell structure. To expand the inventory of potentially relevant proteins, we expressed Ecadherin fused to biotin ligase in MDCK epithelial cells, and identified by mass spectrometry neighboring proteins that were biotinylated. The most abundant of the 303 proteins identified were catenins and nearly 40 others that had been previously reported to influence cadherin function. Many others could be rationalized as novel candidates for regulating the adherens junction, cytoskeleton, trafficking or signaling. We further characterized lipoma preferred partner (LPP), which is present at both cell contacts and focal adhesions. Knockdown of LPP demonstrated its requirement for Ecadherin-dependent adhesion and suggested that it plays a role in coordination of the cell-cell and cell-substrate cytoskeletal interactions. The analysis of LPP function demonstrates proof of principle that the proteomic analysis of E-cadherin proximal proteins expands the inventory of components and tools for understanding the function of E-cadherin.

show abstract

Section: Introductionsupporting

confidence: 72%

Section: Ecadbl Fusion Protein Localizes To Cell-cell Contacts In MDCcontrasting

confidence: 46%

Section: Ecadbl Fusion Protein Localizes To Cell-cell Contacts In MDCmentioning

confidence: 99%

See 1 more Smart Citation

Biotin ligase tagging identifies proteins proximal to E-cadherin, including lipoma preferred partner, a regulator of epithelial cell-cell and cell-substrate adhesion

Itallie

Tietgens

Aponte

et al. 2013

Journal of Cell Science

Self Cite

116

View full text Add to dashboard Cite

show abstract

“…Previous application of BioID to lamin A (LaA) suggested that a majority of the candidates resided within ∼20-30 nm of nuclear envelope (NE)-associated LaA (1). Significantly, distinct subsets of BioID candidates were identified when BirA* was fused to the N versus the C terminus of the cell-junction protein ZO-1 (3). These studies suggested that BioD has a limited nanometer-scale (<20 nm) labeling radius.…”

mentioning

confidence: 87%

“…Issues related to bait (and prey) protein solubility and the stability and/or duration of their interaction are thus overcome. BioID has been used successfully to screen for constituents of the relatively insoluble mammalian nuclear lamina (1), the trypanosome bilobe (2), cell junction complexes (3)(4)(5), and centrosomes (6,7). The method also has been used to screen for proteins involved in the Hippo signaling pathway (8).…”

mentioning

confidence: 99%

Probing nuclear pore complex architecture with proximity-dependent biotinylation

Kim

Zhu

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

464

459

View full text Add to dashboard Cite

Proximity-dependent biotin identification (BioID) is a method for identifying protein associations that occur in vivo. By fusing a promiscuous biotin ligase to a protein of interest expressed in living cells, BioID permits the labeling of proximate proteins during a defined labeling period. In this study we used BioID to study the human nuclear pore complex (NPC), one of the largest macromolecular assemblies in eukaryotes. Anchored within the nuclear envelope, NPCs mediate the nucleocytoplasmic trafficking of numerous cellular components. We applied BioID to constituents of the Nup107-160 complex and the Nup93 complex, two conserved NPC subcomplexes. A strikingly different set of NPC constituents was detected depending on the position of these BioID-fusion proteins within the NPC. By applying BioID to several constituents located throughout the extremely stable Nup107-160 subcomplex, we refined our understanding of this highly conserved subcomplex, in part by demonstrating a direct interaction of Nup43 with Nup85. Furthermore, by using the extremely stable Nup107-160 structure as a molecular ruler, we defined the practical labeling radius of BioID. These studies further our understanding of human NPC organization and demonstrate that BioID is a valuable tool for exploring the constituency and organization of large protein assemblies in living cells.

show abstract

BioID: A Screen for Protein‐Protein Interactions

2013

View full text Add to dashboard Cite

BioID is a unique method to screen for physiologically relevant protein interactions that occur in living cells. This technique harnesses a promiscuous biotin ligase to biotinylate proteins based on proximity. The ligase is fused to a protein of interest and expressed in cells, where it biotinylates proximal endogenous proteins. Because it is a rare protein modification in nature, biotinylation of these endogenous proteins by BioID fusion proteins enables their selective isolation and identification with standard biotin-affinity capture. Proteins identified by BioID are candidate interactors for the protein of interest. BioID can be applied to insoluble proteins, can identify weak and/or transient interactions, and is amenable to temporal regulation. Initially applied to mammalian cells, BioID has potential application in a variety of cell types from diverse species.

show abstract

The N and C Termini of ZO-1 Are Surrounded by Distinct Proteins and Functional Protein Networks

Cited by 108 publications

References 95 publications

Biotin ligase tagging identifies proteins proximal to E-cadherin, including lipoma preferred partner, a regulator of epithelial cell-cell and cell-substrate adhesion

Biotin ligase tagging identifies proteins proximal to E-cadherin, including lipoma preferred partner, a regulator of epithelial cell-cell and cell-substrate adhesion

Probing nuclear pore complex architecture with proximity-dependent biotinylation

BioID: A Screen for Protein‐Protein Interactions

Contact Info

Product

Resources

About