SAK, asl Physical Interaction report

peptide massfingerprinting

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

SAK

Plk4

bait

Protein A tag

asl

Asterless

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 cell extract of S2 cell line; bait produced from transfected construct; prey produced from endogenous gene.

(Dzhindzhev et al., 2010)

FBrf0212014-4.coIP.WB

Reference

Dzhindzhev et al., 2010

Description

physical association

Assay

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

SAK

Plk4

bait

Protein A tag

asl

Asl

prey

GFP tag

asl

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

asl

N-terminal region

necessary binding region

aa 24-55

construct "M1", coordinates relative to asl-PA

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Source was cell extract of S2 cell line; bait produced from transfected construct; prey produced from endogenous gene.

(Dzhindzhev et al., 2010)

FBrf0212014-5.PD.A

Reference

Dzhindzhev et al., 2010

Description

physical association

Assay

autoradiography

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

SAK

Plk4

bait

GST tag

asl

Asl

prey

[35]S label

asl

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

asl

N-terminal region

sufficient binding region

aa 1-300

coordinates relative to asl-PA

asl

N-terminal region

necessary binding region

aa 24-55

construct "M1", coordinates relative to asl-PA

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.

(Dzhindzhev et al., 2010)

FBrf0212014-6.PD.MW

Reference

Dzhindzhev et al., 2010

Description

physical association

Assay

molecular weight estimation by staining

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

asl

Asl

bait

GST tag

asl

SAK

Plk4

prey

MBP tag

SAK

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

asl

N-terminal region

sufficient binding region

aa 1-400

coordinates relative to asl-PA

asl

N-terminal region

necessary binding region

aa 24-55

construct "M1", coordinates relative to asl-PA

SAK

cryptic polo box

sufficient binding region

aa 376-525

coordinates relative to SAK-PA

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 as a recombinant fusion protein in bacterial system.

(Dzhindzhev et al., 2010)

FBrf0212014-7.PD.A

Reference

Dzhindzhev et al., 2010

Description

physical association

Assay

autoradiography

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

asl

Asl

bait

GST tag

asl

SAK

Plk4

prey

[35]S label

SAK

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

asl

N-terminal region

sufficient binding region

aa 1-400

coordinates relative to asl-PA

SAK

cryptic polo box

sufficient binding region

aa 376-525

coordinates relative to SAK-PA

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.

(Dzhindzhev et al., 2010)

FBrf0223435-1.coIP.WB

Reference

Klebba et al., 2013

Description

physical association

Assay

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

SAK

Plk4

bait

GFP tag

SAK

asl

Asl

prey

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

SAK

PB1-PB2

necessary binding region

aa 382-602

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Source was cell extract of S2 cell line; bait produced from transfected construct; prey produced from endogenous gene.

(Klebba et al., 2013)

asl interacts with wild-type or kinase-dead SAK.

(Klebba et al., 2013)

FBrf0223435-5.coIP.MS

Reference

Klebba et al., 2013

Description

physical association

Assay

peptide massfingerprinting

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

SAK

Plk4

bait

Protein A tag

asl

Asl

prey

23 unique peptides identified

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 cell extract of S2 cell line; bait produced from transfected construct; prey produced from endogenous gene.

(Klebba et al., 2013)

asl interacts with wild-type or kinase-dead SAK.

(Klebba et al., 2013)

FBrf0225861-2.PD.WB

Reference

Shimanovskaya et al., 2014

Description

physical association

Assay

microscale thermophoresis

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

SAK

Plk4

bait

His tag

SAK

asl

Asl

prey

MBP tag

asl

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

SAK

cryptic polo box domain

sufficient binding region

aa 383-601

asl

acidic region

sufficient binding region

aa 21-60

SAK

dimerization region residue 490

mutation disrupting interaction

aa 490

R490E

SAK

dimerization region residue 590-591

mutation disrupting interaction

aa 590-591

R594E, R595E

SAK

dimerization region residue 498

mutation decreasing interaction

aa 498

K498E

SAK

dimerization region residue 510

mutation decreasing interaction

aa 510

K510E

SAK

dimerization region residue 523

mutation decreasing interaction

aa 523

R523E

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 as a recombinant fusion protein in bacterial system.

(Shimanovskaya et al., 2014)

FBrf0225861-3.MST

Reference

Shimanovskaya et al., 2014

Description

physical association

Assay

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

SAK

Plk4

bait

asl

Asl

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

Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced as a recombinant fusion protein in bacterial system.

(Shimanovskaya et al., 2014)

The SAK CPB domain binds the asl acidic region with Kd = 0.88 uM.

(Shimanovskaya et al., 2014)

FBrf0225861-4.SLS

Reference

Shimanovskaya et al., 2014

Description

physical association

Assay

static light scattering

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

SAK

Plk4

neutral component

SAK

Plk4

neutral component

SAK

Plk4

neutral component

asl

Asl

neutral component

asl

Asl

neutral component

asl

Asl

neutral component

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

Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey produced as a recombinant fusion protein in bacterial system.

(Shimanovskaya et al., 2014)

FBrf0225974-4.PD.MS

Reference

Cunha-Ferreira et al., 2013

Description

physical association

Assay

peptide massfingerprinting

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

SAK

PLK4

bait

Protein A tag

asl

Asterless

prey

23 unique peptides identified

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 cell extract of S2 cell line; bait produced from transfected construct; prey produced from endogenous gene.

(Cunha-Ferreira et al., 2013)

FBrf0226888-3.ENZ.A

Reference

Description

Assay

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

SAK

Plk4

enzyme

MBP tag

SAK

Plk4

enzyme

MBP tag

asl

Asl

enzyme target

GST tag, P[32] label

asl

Asl

enzyme target

GST tag, P[32] label

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

FBrf0227526-11.Y2H

Reference

Klebba et al., 2015

Description

physical association

Assay

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

asl

Asl

bait

Asl-C fragment

asl

SAK

Plk4

prey

-PB2 domain fragment

SAK

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

asl

Asl-C domain

sufficient binding region

aa 626-994

asl

Asl-C domain mutation 1

mutation disrupting interaction

E698G

asl

Asl-C domain mutation 2

mutation disrupting interaction

E789D

asl

Asl-C domain mutation 3

mutation disrupting interaction

S906P

asl

Asl-C domain mutation 4

mutation disrupting interaction

M832V

asl

Asl-C domain mutation 5

mutation disrupting interaction

M872T

asl

Asl-C domain mutation 6

mutation disrupting interaction

Q884L

asl

Asl-C domain mutation 7

mutation disrupting interaction

E926G

asl

Asl-C domain mutation 8

mutation disrupting interaction

K667E

asl

Asl-C domain mutation 9

mutation disrupting interaction

E901G

asl

Asl-C domain mutation 10

mutation disrupting interaction

Y921H

asl

Asl-C domain mutation 11

mutation disrupting interaction

I648T

asl

Asl-C domain mutation 12

mutation disrupting interaction

K731R

asl

Asl-C domain mutation 13

mutation disrupting interaction

L840P

asl

Asl-C domain mutation 14

mutation disrupting interaction

R910stop

SAK

PB1 domain

necessary binding region

Polo box

SAK

PB2 domain

necessary binding region

Polo box PB2

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.

(Klebba et al., 2015)

Extensive set of amino acid substitutions for asl across five yeast two-hybrid constructs.

(Klebba et al., 2015)

FBrf0227526-2.coIP.WB

Reference

Klebba et al., 2015

Description

physical association

Assay

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

asl

Asl

bait

V5 tag

asl

SAK

Plk4

prey

EGFP tag

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

asl

Asl-C domain

sufficient binding region

aa 626-994

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Source was cell extract of S2 cell line; bait produced from transfected construct; prey produced from transfected construct.

(Klebba et al., 2015)

Authors indicate asl interacts with SAK primarily through its C terminus.

(Klebba et al., 2015)

FBrf0227526-3.coIP.WB

Reference

Klebba et al., 2015

Description

physical association

Assay

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

SAK

Pkl4

bait

EGFP tag

SAK

asl

Asl

prey

V5 tag, C fragment

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

SAK

PB1 domain

necessary binding region

Polo box PB1

SAK

PB2 domain

necessary binding region

Polo box PB2

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Interaction requires the presence of Polo boxes PB1 and PB2 for SAK.

(Klebba et al., 2015)

Source was cell extract of S2 cell line; bait produced from transfected construct; prey produced from transfected construct.

(Klebba et al., 2015)

The combined PB1 and PB2 domains of SAK span amino acids 382-602.

(Klebba et al., 2015)

FBrf0227526-4.Y2H

Reference

Klebba et al., 2015

Description

physical association

Assay

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

SAK

Pkl4

bait

SAK

asl

Asl

prey

asl

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

SAK

PB1 domain

necessary binding region

Polo box PB1

SAK

PB2 domain

necessary binding region

Polo box PB2

asl

Asl-C domain

sufficient binding region

aa 626-994

Asl-C domain

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

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

(Klebba et al., 2015)

Source was yeast cell line; bait produced as transgenic fusion protein; prey produced as transgenic fusion protein.

(Klebba et al., 2015)

The combined PB1 and PB2 domains of SAK span amino acids 382-602.

(Klebba et al., 2015)

FBrf0228377-3.coIP.WB

Reference

Klebba et al., 2015

Description

physical association

Assay

Collection

Source/Stage

Cell line used

S2-unspecified

Participants

Corresponds to

Reported as

Role

Note

SAK

bait

GFP tag

SAK

asl

prey

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

SAK

PB1 domain

necessary binding region

aa 381-500

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Source was cell extract of S2 cell line; bait produced from transfected construct; prey produced from endogenous gene.

(Klebba et al., 2015)

FBrf0232405-4.Y2H

Reference

Galletta et al., 2016

Description

physical association

Assay

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

asl

Asl

bait

fused to GAL4 DNA-binding domain

SAK

Plk4

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).

(Galletta et al., 2016)

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

(Galletta et al., 2016)

FBrf0233282-63.Y2H

Reference

Galletta et al., 2016

Description

physical association

Assay

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

asl

Asl

bait

fused to GAL4 DNA-binding domain

asl

SAK

Plk4/SAK

prey

fused to GAL4 activation domain

SAK

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

asl

N-terminal region

sufficient binding region

aa 1-357

coordinates relative to asl-PA

SAK

central region

sufficient binding region

aa 382-602

coordinates relative to SAK-PA

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

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

(Galletta et al., 2016)

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

(Galletta et al., 2016)

FBrf0233282-64.Y2H

Reference

Galletta et al., 2016

Description

physical association

Assay

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

SAK

Plk4/SAK

bait

fused to GAL4 DNA-binding domain

SAK

asl

Asl

prey

fused to GAL4 activation domain

asl

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

asl

C-terminal region

sufficient binding region

aa 626-994

coordinates relative to asl-PA

SAK

central region

sufficient binding region

aa 382-602

coordinates relative to SAK-PA

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

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

(Galletta et al., 2016)

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

(Galletta et al., 2016)

FBrf0240774-1.ENZ.A

Reference

Description

Assay

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

SAK

Plk4

enzyme

His tag

SAK

Plk4

enzyme

His tag

SAK

asl

Asl

enzyme target

GST tag, P[32] label

asl

Asl

enzyme target

GST tag, P[32] label

asl

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

FBrf0240774-5.coIP.WB

Reference

Boese et al., 2018

Description

physical association

Assay

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

asl

Asl

bait

GFP tag

SAK

Plk4

prey

Myc tag

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 cell extract of S2 cell line; bait produced from transfected construct; prey produced from transfected construct.

(Boese et al., 2018)

FBrf0240774-6.coIP.WB

Reference

Boese et al., 2018

Description

physical association

Assay

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

SAK

Plk4

bait

GFP tag

SAK

asl

Asl

prey

V5 tag

asl

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

SAK

PB1-PB2 domain

sufficient binding region

aa 382-602

SAK

PB1-PB2 domain

necessary binding region

aa 382-602

asl

N-terminal domain

sufficient binding region

aa 1-374

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Source was cell extract of S2 cell line; bait produced from transfected construct; prey produced from transfected construct.

(Boese et al., 2018)

FBrf0246771-10.coIP.WB

Reference

McLamarrah et al., 2020

Description

physical association

Assay

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

asl

Asl

bait

GFP tag

asl

SAK

Plk4

prey

His tag

SAK

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

SAK

interaction domain

sufficient binding region

aa 381-602

coordinates relative to SAK-PA

asl

interaction domain

sufficient binding region

aa 1-357

coordinates relative to asl-PA

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Source was bacterial lysate; proteins coproduced as a recombinant fusion proteins.

(McLamarrah et al., 2020)

FBrf0246771-4.PD.WB

Reference

McLamarrah et al., 2020

Description

physical association

Assay

Collection

Source/Stage

Cell line used

Participants

Corresponds to

Reported as

Role

Note

SAK

Plk4

bait

His tag

SAK

asl

Asl

prey

GFP tag

asl

Experimental entities

Corresponds to

Identifier

Reported

Role

Note

Subregions with role in interaction

Corresponds to

Description

Role

Coordinates

Note

SAK

interaction domain

sufficient binding region

aa 381-602

coordinates relative to SAK-PA

asl

interaction domain

sufficient binding region

aa 1-357

coordinates relative to asl-PA

Isoform-specific participants

Corresponds to

Description

Role

Note

Comments concerning this interaction

Source was bacterial lysate; proteins coproduced as a recombinant fusion proteins.

(McLamarrah et al., 2020)

FBrf0255743-8.PD.A

Reference

Ábrahám et al., 2023

Description

physical association

Assay