insc, pins Physical Interaction report

Collection

Source/Stage

organism / embryonic stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

insc

Insc

bait

FLAG tag

pins

Pins

prey

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Source was embryos of transgenic fly line; bait produced from tagged transgenic construct; prey produced from endogenous gene.

(Yu et al., 2000)

FBrf0125333-4.PD.A

Reference

Yu et al., 2000

Description

physical association

Assay

anti tag coimmunoprecipitation

autoradiography

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

insc

Insc

bait

GST tag

insc

pins

Pins

prey

[35]S label

pins

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

insc

asymmetric localization domain

sufficient binding region

aa 288-497

pins

N-terminal region

sufficient binding region

aa 1-378

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced and labeled by in vitro translation.

(Yu et al., 2000)

FBrf0127308-1.coIP.MS

Reference

Schaefer et al., 2000

Description

physical association

Assay

peptide massfingerprinting

Collection

Source/Stage

organism / embryonic stage 10 embryonic stage 12 embryonic stage 11

Cell line used

Participants

Corresponds to

Reported as

Role

Note

insc

Insc

bait

β-gal tag

insc

pins

Pins

prey

6 unique peptides identified

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

insc

membrane localisation domain

sufficient binding region

aa 302-459

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Source was embryos of transgenic fly line; bait produced from tagged transgenic construct; prey produced from endogenous gene.

(Schaefer et al., 2000)

FBrf0127308-4.coIP.WB

Reference

Schaefer et al., 2000

Description

physical association

Assay

anti tag coimmunoprecipitation

Collection

Source/Stage

organism / embryonic stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

insc

Insc

bait

β-gal tag

insc

pins

Pins

prey

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

insc

central asymmetry domain

sufficient binding region

aa 252-615

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Source was embryos of transgenic fly line; bait produced from tagged transgenic construct; prey produced from endogenous gene.

(Schaefer et al., 2000)

FBrf0127308-5.PD.A

Reference

Schaefer et al., 2000

Description

physical association

Assay

autoradiography

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

insc

Insc

bait

GST tag

insc

pins

Pins

prey

[35]S label

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

insc

central asymmetry domain

sufficient binding region

aa 252-615

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced and labeled by in vitro translation.

(Schaefer et al., 2000)

FBrf0217064-1.XC

Reference

Culurgioni et al., 2011

Description

physical association

Assay

x-ray crystallography

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

pins

Pins

unspecified role

pins

Pins

unspecified role

pins

Pins

unspecified role

pins

insc

Insc

unspecified role

insc

Insc

unspecified role

insc

Insc

unspecified role

insc

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

pins

region

sufficient binding region

aa 39-386

\'TPR\'; includes TPR repeats

insc

region

sufficient binding region

aa 307-335

\'PEPT\'

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Cloned protein fragments coexpressed in bacteria via a dicistronic vector; purified as a complex by size exclusion chromatography.

(Culurgioni et al., 2011)

FBrf0217064-3.PD.gel

Reference

Culurgioni et al., 2011

Description

physical association

Assay

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

insc

Insc

bait

GST pull down

insc

pins

Pins

prey

pins

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

pins

region

sufficient binding region

aa 25-406

\'TPR\'; includes TPR repeats

insc

region

sufficient binding region

aa 303-340

\'PEPT\'

pins

region

necessary binding region

aa 226

Removal of side chain of N226 eliminates binding.

pins

RR258/259AA

necessary binding region

aa 258-259

Replacement of R258 and R259 with Ala eliminates binding.

insc

EE322/324AA

necessary binding region

aa 322-324

Replacement of E322 and E324 with Ala eliminates binding.

insc

EE322/324RR

necessary binding region

aa 322-324

Replacement of E322 and E324 with Arg eliminates binding.

insc

W313A

necessary binding region

aa 313

Replacement of W313 with Ala eliminates binding.

insc

W313E

necessary binding region

aa 313

Replacement of W313 with Glu eliminates binding.

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Cloned reagents; interaction in solution.

(Culurgioni et al., 2011)

FBrf0218437-1.PD.MW

Reference

Mauser and Prehoda, 2012

Description

physical association

Assay

anti bait coimmunoprecipitation

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

pins

Pins

bait

GST tag

pins

insc

Insc

prey

MBP tag

insc

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

insc

ankyrin repeat domain

sufficient binding region

aa 252-600

pins

TPR domain

sufficient binding region

aa 42-398

pins

TPR repeats 4-7

necessary binding region

deletion greatly weakens interaction

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

GST pull down.

(Mauser and Prehoda, 2012)

Bait expressed and purified from bacteria.

(Mauser and Prehoda, 2012)

Prey expressed and purified from bacteria.

(Mauser and Prehoda, 2012)

All 7 tetratricopeptide repeat (TPR) motifs of raps are required for high affinity binding to insc.

(Mauser and Prehoda, 2012)

FBrf0218437-2.coIP.WB

Reference

Mauser and Prehoda, 2012

Description

physical association

Assay

anti bait coimmunoprecipitation

Collection

Source/Stage

larval brain / third instar larval stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

insc

Insc

bait

pins

Pins

prey

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Given that raps coimmunoprecipitates with insc and mud, but insc and mud do not coimmunoprecitate each other, the authors conclude that raps forms mutually exclusive complexes with insc and mud.

(Mauser and Prehoda, 2012)

Bait endogenous to larval brain extract.

(Mauser and Prehoda, 2012)

Prey endogenous to larval brain extract.

(Mauser and Prehoda, 2012)

FBrf0218437-4.coIP.WB

Reference

Mauser and Prehoda, 2012

Description

physical association

Assay

Collection

Source/Stage

larval brain / third instar larval stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

pins

Pins

bait

insc

Insc

prey

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Given that raps coimmunoprecipitates with insc and mud, but insc and mud do not coimmunoprecitate each other, the authors conclude that raps forms mutually exclusive complexes with insc and mud.

(Mauser and Prehoda, 2012)

Bait endogenous to larval brain extract.

(Mauser and Prehoda, 2012)

Prey endogenous to larval brain extract.

(Mauser and Prehoda, 2012)

FBrf0218437-8.PD.MW

Reference

Mauser and Prehoda, 2012

Description

physical association

Assay

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

pins

Pins

bait

GST tag

pins

insc

Insc

prey

MBP tag

insc

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

pins

TPR domain

sufficient binding region

aa 42-398

insc

ankyrin repeat domain

sufficient binding region

aa 252-600

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

GST pull down.

(Mauser and Prehoda, 2012)

Bait expressed and purified from bacteria.

(Mauser and Prehoda, 2012)

Prey expressed and purified from bacteria.

(Mauser and Prehoda, 2012)

mud and insc compete for binding to raps.

(Mauser and Prehoda, 2012)

FBrf0220918-19.Y2H

Reference

Januschke et al., 2013

Description

physical association

Assay

two hybrid

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

insc

Insc

bait

fused to GAL4 DNA-binding domain

pins

Pins

prey

fused to GAL4 activation domain

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein (prey was previously cloned reagent).

(Januschke et al., 2013)

Two-hybrid system: yeast GAL4-BD/GAL4-AD

(Januschke et al., 2013)

Positive control.

(Januschke et al., 2013)

FBrf0238321-1.XC

Reference

Culurgioni et al., 2018

Description

physical association

Assay

x-ray crystallography

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

pins

dLGN

neutral component

pins

dLGN

neutral component

pins

dLGN

neutral component

pins

insc

Insc

neutral component

insc

Insc

neutral component

insc

Insc

neutral component

insc

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

pins

TPR domain

sufficient binding region

aa 25-406

insc

ASYM domain

sufficient binding region

aa 283-623

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Structure is a heterotetramer with 2:2 stoichiometry.

(Culurgioni et al., 2018)

Interaction in vitro; proteins produced as recombinant proteins in bacterial system.

(Culurgioni et al., 2018)

FBrf0238321-2.SLS.MW

Reference

Culurgioni et al., 2018

Description

physical association

Assay

static light scattering

molecular weight

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

pins

dLGN

neutral component

pins

dLGN

neutral component

pins

dLGN

neutral component

pins

insc

Insc

neutral component

insc

Insc

neutral component

insc

Insc

neutral component

insc

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

pins

TPR domain

sufficient binding region

aa 25-406

insc

ASYM domain

sufficient binding region

aa 283-623

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Molecular mass estimated by SLS is consistent with a 2:2 heterotetramer.

(Culurgioni et al., 2018)

Interaction in vitro; proteins produced as recombinant proteins in bacterial system.

(Culurgioni et al., 2018)

FBrf0238321-3.CH.MW

Reference

Culurgioni et al., 2018

Description

physical association

Assay

molecular sieving

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

pins

dLGN

neutral component

pins

dLGN

neutral component

pins

dLGN

neutral component

pins

insc

Insc

neutral component

insc

Insc

neutral component

insc

Insc

neutral component

insc

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

pins

TPR domain

sufficient binding region

aa 25-406

insc

ASYM domain

sufficient binding region

aa 283-623

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Interaction in vitro; proteins produced as a recombinant fusion proteins in bacterial system.

(Culurgioni et al., 2018)

FBrf0257436-4.PD.MW

Reference

Parra et al., 2023

Description

physical association

Assay