================================================
note: expressed and recombinant Irak1 are active
================================================
Irak1 kinase activity
================================================

requires D340 (active site)

  *** xIrak1[tAb]IP IVKA(auto)[32P-ATP] is detectable
    *** cells: none
    *** IPfrom: SF9 in BMS
    *** inhibited by: xIrak1(D340N)"KD"[tAb]IP [substitution]
    *** source: 10191251-Fig-1b

requires K239 (ATP binding site)

  *** rIrak1[32P] IVKA(auto)[32P-ATP] is detectable
    *** cells: none
    *** inhibited by: xIrak1(K239S)"KD" [substitution]
    *** source: 12860405-Fig-2a
  *** rIrak1[32P] phos[32P-ATP] is detectable
    *** cells: none
    *** inhibited by: rIrak1(K239S)"KD" [substitution]
    *** source: 11960013-Fig-5a
  *** xIrak1[Ab]IP IVKA(auto)[32P-ATP] is detectable
    *** cells: HEK293<Irak1~null> in BMS
    *** inhibited by: xIrak1(K239A)"KD" [substitution]
    *** source: 10373513-Fig-8
  *** xIrak1[tAb]IP IVKA(auto)[32P-ATP] is detectable
    *** cells: HEK293 in BMS
    *** inhibited by: xIrak1(K239A)"KD" [substitution]
    *** source: 15767370-Fig-2b
  *** xIrak1[tAb]IP IVKA(auto)[32P-ATP] is detectable
    *** cells: HEK293 in BMS
    *** inhibited by: xIrak1(K239S) [substitution]
    *** source: 14625308-Fig-2b
  *** xIrak1[tAb]IP IVKA(auto)[32P-ATP] is detectable
    *** cells: HEK293 in BMS
    *** inhibited by: xIrak1(K239S) [substitution]
    *** source: 14625308-Fig-4a

requires T209 (phos site)

  *** xIrak1[tAb]IP IVKA(auto)[32P-ATP] is detectable
    *** cells: HEK293 in BMS
    *** inhibited by: xIrak1(T209A) [substitution]
    *** inhibited by: xIrak1(T209D) [substitution]
    *** inhibited by: xIrak1(T209E) [substitution]
    *** source: 14625308-Fig-2b

partially requires T387 (activation loop)

  *** xIrak1[tAb]IP IVKA(auto)[32P-ATP] is detectable
    *** cells: HEK293 in BMS
    *** partially inhibited by: xIrak1(T387A) [substitution]
    *** source: 14625308-Fig-2b
  *** xIrak1(T387A)[tAb]IP IVKA(auto)[32P-ATP] is detectable
    *** cells: HEK293 in BMS
    *** unaffected by: xIrak1(T387A/S365A) [substitution]
    *** unaffected by: xIrak1(T387A/S307A) [substitution]
    *** unaffected by: xIrak1(T387A/S371A) [substitution]
    *** unaffected by: xIrak1(T387A/S373A) [substitution]
    *** unaffected by: xIrak1(T387A/S375A) [substitution]
    *** unaffected by: xIrak1(T387A/S376A) [substitution]
    *** unaffected by: xIrak1(T387A/S381A) [substitution]
    *** unaffected by: xIrak1(T387A/S383A) [substitution]
    *** source: 14625308-Fig-4b
  *** xIrak1[tAb]IP IVKA(auto)[32P-ATP] is detectable
    *** cells: HEK293 in BMS
    *** partially inhibited by: xIrak1(T387A) [substitution]
    *** source: 14625308-Fig-4a

does not req S365, S370, S371, S373, S375, S376, S381, S383 (kinase domain)
  *** xIrak1[tAb]IP IVKA(auto)[32P-ATP] is detectable
    *** cells: HEK293 in BMS
    *** unaffected by: xIrak1(S365A) [substitution]
    *** enhanced by: xIrak1(S370A) [substitution]
    *** enhanced by: xIrak1(S371A) [substitution]
    *** enhanced by: xIrak1(S373A) [substitution]
    *** enhanced by: xIrak1(S375A) [substitution]
    *** enhanced by: xIrak1(S376A) [substitution]
    *** enhanced by: xIrak1(S381A) [substitution]
    *** unaffected by: xIrak1(S383A) [substitution]
    *** source: 14625308-Fig-4a

does not req phos on T66 ???

  *** xIrak1[tAb]IP IVKA(auto)[32P-ATP] is detectable
    *** cells: HEK293<xIL1R1> in BMS
    *** inhibited by: xIrak1(K239S) [substitution]
    *** unaffected by: xIrak1(T66A) [substitution]
    *** inhibited by: xIrak1(T66D) [substitution]
    *** inhibited by: xIrak1(T66E) [substitution]
    *** unaffected by: xIrak1(T66N) [substitution]
    *** source: 18524972-Fig-4

does not req phos in ST reqion

  *** xIrak1[tAb]IP IVKA(auto)[32P-ATP] is detectable
    *** cells: HEK293<xIL1R1> in BMS
    *** unaffected by: xIrak1(del(110-211)) [substitution]
    *** source: 18524972-Fig-6

  *** Irak1[Ab]IP IVKA(auto)[32P-ATP] is detectable
    *** cells: HEK293 in BMS
    *** source: 10373513-Fig-8

=======================================================================
Evidence that the large upshift in mobibility is due to phosphorylation
=======================================================================

  *** xIrak1[Ab] upshift[WBMS] is detectable
    *** cells: HEK293 in BMS
    *** comment: inhibited by treatment of lysates with PP2a
    *** inhibited by: xIrak1(D340N) [substitution]
    *** source: 10191251-Fig-1a
  *** xIrak1[Ab] upshift[WBMS] is detectable
    *** cells: HEK293T in BMS
    *** comment: inhibited by treatment of lysates with PPase
    *** source: 10854325-Fig-3e
  *** xIrak1(T66A)[Ab] upshift[WBMS] is detectable
    *** cells: HEK293T in BMS
    *** comment: inhibited by treatment of lysates with PPase
    *** source: 10854325-Fig-3e

========================
Irak1 autophophorylation
========================

on T387/ S376

  *** sIrak1(359-389)[32P] phos[32P-ATP] is increased by rIrak1
    *** cells: none
    *** partially inhibited by: sIrak1((359-389)/pT387) [substitution]
    *** inhibited by: sIrak1((359-389)/pS376) [substitution]
    *** source: 11960013-Fig-5b

on S137/S138

  *** xIrak1(110-211)[tAb]IP phos[32P-ATP] is increased by xIrak1[tAb]IP
    *** cells: none
    *** comment: IPs are from HEK293 in BMS containing both constructs
    *** unaffected by: xIrak1(110-208) [substitution]
    *** unaffected by: xIrak1(110-195) [substitution]
    *** unaffected by: xIrak1(110-184) [substitution]
    *** unaffected by: xIrak1(110-176) [substitution]
    *** unaffected by: xIrak1(110-168) [substitution]
    *** unaffected by: xIrak1(110-155) [substitution]
    *** unaffected by: xIrak1(110-151) [substitution]
    *** unaffected by: xIrak1(110-143) [substitution]
    *** unaffected by: xIrak1(110-139) [substitution]
    *** inhibited by: xIrak1(110-136) [substitution]
    *** inhibited by: xIrak1(110-127) [substitution]
    *** source: 14625308-Fig-5b

on T209

  *** xIrak1(149-536)[tAb]IP phos[32P-ATP] is detectable
    *** cells: HEK293 in BMS
    *** unaffected by: xIrak1(194-536) [substitution]
    *** partially inhibited by: xIrak1((194-536)/S196A) [substitution]
    *** partially inhibited by: xIrak1((194-536)/S206A) [substitution]
    *** inhibited by: xIrak1((194-536)/T209A) [substitution]
    *** inhibited by: xIrak1(212-536) [substitution]
    *** source: 14625308-Fig-2a

not T66

  *** xIrak1[tAb] upshift[WBMS] is detectable
    *** cells: HEK293<xIL1R1> in BMS
    *** inhibited by: xIrak1(K239S) [substitution]
    *** unaffected by: xIrak1(T66A) [substitution]
    *** partially inhibited by: xIrak1(T66D) [substitution]
    *** inhibited by: xIrak1(T66E) [substitution]
    *** unaffected by: xIrak1(T66N) [substitution]
    *** source: 18524972-Fig-4




  *** xIrak1(2-208)[tAb]IP phos[32P-ATP] is increased by xIrak1[tAb]IP
    *** cells: none
    *** comment: IPs are from HEK293 in BMS containing both constructs
    *** source: 14625308-Fig-1c
  *** xIrak1(149-536)[tAb]IP phos[32P-ATP] is detectable
    *** cells: HEK293 in BMS
    *** source: 14625308-Fig-1c
  *** xIrak1(212-536)[tAb]IP phos[32P-ATP] is unchanged by xIrak1[tAb]IP
    *** cells: none
    *** comment: IPs are from HEK293 in BMS containing both constructs
    *** unaffected by: xIrak1((212-536)/K239S) [substitution]
    *** source: 14625308-Fig-1c

===============
oligomerization
===============

  *** rIrak1[Ab] oligomerization[density-gradient] is detectable
    *** cells: none
    *** inhibited by: rIrak1(T66E) [substitution]
    *** source: 16831874-Fig-7a
  *** rIrak1b[Ab] oligomerization[density-gradient] is detectable
    *** cells: none
    *** source: 16831874-Fig-7a

requires T387 (phos site in activation loop)
  *** xIrak1[tAb]IP upshift[WBMS] is detectable
    *** cells: HEK293 in BMS
    *** comment: mobililty shift seen in xIrak1 is not seen in xIrak1(T387A)
    *** source: 14625308-Fig-2b,4b
  *** xIrak1[tAb]IP upshift[WBMS] is detectable
    *** cells: HEK293 in BMS
    *** comment: mobililty shift seen in xIrak1 is not seen in xIrak1(T387A)
    *** source: 14625308-Fig-4a

requires D340N (active site)
  *** xIrak1[Ab] upshift[WBMS] is detectable
    *** cells: HEK293T in BMS
    *** inhibited by: xIrak1(D340N)"KD" [substitution]
    *** source: 10854325-Fig-3c

partially requires T66 (death domain phos site)
  *** xIrak1[Ab] upshift[WBMS] is detectable
    *** cells: HEK293T in BMS
    *** partially inhibited by: xIrak1(T66A) [substitution]
    *** source: 10854325-Fig-3c


==============
Sumoylation??
==============

  *** Irak1[Ab]IP sumo[SuAb] is detectable
    *** cells: THP1 in BMS
    *** unaffected by: Lps (1 hr) [stim]
    *** unaffected by: Pam3Csk4 (1 hr) [stim]
    *** source: 16690127-Fig-2a
  *** xIrak1[tAb]IP sumo[SuAb] is detectable
    *** cells: THP1 in BMS
    *** comment: most of the sumo-Irak1 is in the nuclear-fraction
    *** source: 16690127-Fig-2c

================
coprecipitations
================

Bcl10--
  *** Irak1[Ab] copptby[WB] xBcl10[tAb] is detectable
    *** cells: RAW2647 in BMS "treated with Lps (30 min)"
    *** inhibited by: xBcl10(L41Q/G78R)"oligo-mut" [addition]
    *** source: 16831874-Fig-6c
  *** Irak1[Ab] copptby[WB] Bcl10[Ab] is undetectable
    *** cells: RAW2647 in BMS
    *** source: 16831874-Fig-10c
Btk--
  *** xIrak1[Ab] copptby[WB] xBtk[Ab] is detectable
    *** cells: HEK293 in BMS
    *** source: 12724322(D)
IL1Rap--
  *** Irak1[Ab] copptby[WB] xIL1Rap[tAb] is detectable
    *** cells: HEK293 in BMS
    *** source: 9371760-Fig-3b

Irak4--
  *** xIrak4[Ab] copptby[WB] xIrak1(del(110-211))[tAb] is detectable
    *** cells: HEK293<xIL1R1> in BMS
    *** inhibited by: xIrak1(WT) [substitution]
    *** source: 18524972-Fig-6
  *** xIrak4[Ab] copptby[WB] xIrak1(K239S)[tAb] is detectable
    *** cells: HEK293<xIL1R1> in BMS
    *** inhibited by: xIrak1(WT) [substitution]
    *** unaffected by: xIrak1(T66A) [substitution]
    *** unaffected by: xIrak1(T66D) [substitution]
    *** unaffected by: xIrak1(T66E) [substitution]
    *** unaffected by: xIrak1(T66N) [substitution]
    *** source: 18524972-Fig-1
Malt1--
  *** Irak1[Ab] copptby[WB] Malt1[Ab] is undetectable
    *** cells: RAW2647 in BMS
    *** unaffected by: Lps (30 min) [stim]
    *** source: 16831874-Fig-10c
Myd88--
  *** xIrak1(K239S)"KD"[tAb] copptby[WB] xMyd88[tAb] is detectable
    *** cells: HEK293 in BMS
    *** inhibited by: xIrak1(WT) [substitution]
    *** source: 11960013-Fig-2a
  *** xMyd88[tAb] copptby[WB] xIrak1(K239S)"KD"[Ab] is detectable
    *** cells: HEK293 in BMS
    *** inhibited by: xIrak1(WT) [substitution]
    *** unaffected by: xIrak1(T209A)"KD" [substitution]
    *** unaffected by: xIrak1(del(110-211)) [substitution]
    *** inhibited by: xIrak1(del(212-536)) [substitution]
    *** source: 14625308-Fig-6a
  *** xMyd88[Ab] copptby[WB] xIrak1(del(110-211))[tAb] is detectable
    *** cells: HEK293<xIL1R1> in BMS
    *** inhibited by: xIrak1(WT) [substitution]
    *** source: 18524972-Fig-6
  *** xMyd88[Ab] copptby[WB] xIrak1(K239S)[tAb] is detectable
    *** cells: HEK293<xIL1R1> in BMS
    *** inhibited by: xIrak1(WT) [substitution]
    *** unaffected by: xIrak1(T66A) [substitution]
    *** unaffected by: xIrak1(T66D) [substitution]
    *** unaffected by: xIrak1(T66E) [substitution]
    *** unaffected by: xIrak1(T66N) [substitution]
    *** source: 18524972-Fig-1
  *** xIrak1(K239S)"KD"[Ab] copptby[WB] xMyd88[tAb] is detectable
    *** cells: HEK293 in BMS
    *** inhibited by: xIrak1(WT) [substitution]
    *** source: 9430229-Fig-5a
Peli2--
  *** xIrak1[tAb] copptby[WB] xPeli2[tAb] is detectable
    *** cells: HEK293 in BMS
    *** source: 12860405-Fig-1a
Peli3--
  *** xIrak1[tAb] copptby[WB] xPeli3[tAb] is detectable
    *** cells: HEK293 in BMS
    *** source: 12874243-Fig-5a
Tak1--
  *** Irak1[Ab] copptby[WB] Tak1[Ab] is undetectable
    *** cells: RAW2647 in BMS
    *** source: 16831874-Fig-10c
Tifa--
  *** xIrak1[tAb] snaggedby[WB] rTifa[GST] is detectable
    *** cells: HEK293T in BMS
    *** source: 12566447-Fig-4a
Tollip--
  *** Irak1[Ab] copptby[WB] Tollip[Ab] is detectable
    *** cells: EL4 in BMS
    *** source: 10854325-Fig-3b
  *** xTollip[Ab] copptby[WB] xIrak1(K239S)"KD"[Ab] is detectable
    *** cells: HEK293 in BMS
    *** inhibited by: xIrak1(WT) [substitution]
    *** unaffected by: xIrak1(T209A)"KD" [substitution]
    *** unaffected by: xIrak1(del(110-211)) [substitution]
    *** inhibited by: xIrak1(del(212-536)) [substitution]
    *** source: 14625308-Fig-6a
  *** xIrak1(D340N)"KD"[Ab] copptby[WB] xTollip[tAb] is detectable
    *** cells: HEK293T in BMS
    *** inhibited by: xTollip(del(48-178)) [substitution]
    *** unaffected by: xTollip(47-274) [substitution]
    *** partially inhibited by: xTollip(1-229) [substitution]
    *** inhibited by: xTollip(1-178) [substitution]
    *** source: 10854325-Fig-5b
  *** xIrak1(D340N)"KD"[Ab] copptby[WB] xTollip[tAb] is detectable
    *** cells: HEK293T in BMS
    *** inhibited by: xIrak1(WT) [substitution]
    *** enhanced by: xIrak1(T66A) [substitution]
    *** source: 10854325-Fig-3c
  *** xTollip[Ab] copptby[WB] xIrak1(del(110-211))[tAb] is detectable
    *** cells: HEK293<xIL1R1> in BMS
    *** inhibited by: xIrak1(WT) [substitution]
    *** source: 18524972-Fig-6
  *** xTollip[Ab] copptby[WB] xIrak1(K239S)[tAb] is detectable
    *** cells: HEK293<xIL1R1> in BMS
    *** inhibited by: xIrak1(WT) [substitution]
    *** unaffected by: xIrak1(T66A) [substitution]
    *** unaffected by: xIrak1(T66D) [substitution]
    *** unaffected by: xIrak1(T66E) [substitution]
    *** unaffected by: xIrak1(T66N) [substitution]
    *** source: 18524972-Fig-1
Traf2--
  *** Irak1[Ab] copptby[WB] xTraf2[tAb] is undetectable
    *** cells: HEK293T in BMS
    *** source: 9374458-Fig-3a
Traf3--
  *** xIrak1[tAb] copptby[WB] xTraf3[tAb] is detectable
    *** cells: HEK293T in BMS
    *** source: 16306936-Fig-3d
Traf6--
  *** xIrak1(K239S)"KD"[tAb] copptby[WB] xTraf6[tAb] is detectable
    *** cells: HEK293 in BMS
    *** inhibited by: xIrak1(WT) [substitution]
    *** source: 11960013-Fig-2b
  *** xTraf6[Ab] copptby[WB] xIrak1[Ab] is detectable
    *** cells: HEK293 in BMS
    *** unaffected by: xIrak1(K239S)"KD" [substitution]
    *** unaffected by: xIrak1(T209A)"KD" [substitution]
    *** unaffected by: xIrak1(del(110-211)) [substitution]
    *** unaffected by: xIrak1(del(212-536)) [substitution]
    *** source: 14625308-Fig-6a
  *** Traf6[Ab] copptby[WB] xIrak1[Ab] is detectable
    *** cells: HEK293I1A<Irak1~null> in BMS
    *** inhibited by: xIrak1(K293A)"KD" [substitution]
    *** unaffected by: IL1b (30 min) [stim]
    *** source: 11518704-Fig-7

