TRPC Family

The TRPC Subfamily TRP Subtype Unigene

C1 Hs. 250687

C2 *602343

C3 Hs. 150981

C4 Hs.2 62960

C5 Hs.2 47868

C6 Hs.1 59003

C7 Hs.2 83104

# Amino ac ids ( h)

793 11 72 sev e ral s plice varia nts

848 977 (a ), 893 (b) multiple s plice va riants

973 931 862

OMIM ge ne map (h) (m)

3q22-3q24 9

pse udoge ne 11 p15.3-p15.4

4q27 3

13q13.1-q13.2 Xq23

11 q21-q22 mouse 9

5q31.1 mouse 13

Genetic mo del or dis ease mutation

Mouse TRPC 2-/- Mouse TRPC 4-/- Mouse TRPC 6-/-

Known Func tions

Phe romo ne sens ing in mouse (Vome ronasal organ : VNO)

Vaso regulat ion (Resis tance ar tery myoge nic tone ) Airway re gula tion

Impaired ag onis t-dep e nden t vas orelax at ion in TRPC 4-/- mouse

Growth cone morphology

Vaso regulat ion (Resis tance ar tery myoge nic tone ) Airway re gula tion

Likely role in

Purported me cha no- sensit ive cha nne l, s tretch-sensit ive

CNS Vaso regulat ion, microvascu lar pe rme ability? CNS GI motility 5-HT regula ted GABA relea s e from thala mic inte rneu rons

CNS

Rec e ptor-activa te d role in smoo th musc le, potent ial activation via stre tch s timulat ion of PLCb

N or C terminal do main mo dule s

3x N-AnkR C-TRP box-C-P olyP (EWKFAR…PP P FN) [N+C coi led coils; N+C PKC sites; N PKA site ; TyrK site - C-CIRB; N-Ca v1BD

3x N-AnkR C-TRP box-C-P olyP (EWKFAR…PP P FN) C-CIRB N-PKD1 (TRP P 1) homol ogy

4x N-AnkR C-TRP box-C-P olyP (EWKFAR…PP P FN) C-CIRB

2x N-AnkR C-PDZ binding C-TRP box-C-P olyP (EWKFAR…PP P FN) C-CIRB -C-Cam

2x N-AnkR C-PDZ binding C-TRP box-C-P olyP (EWKFAR…PP P FN) C-CIRB

3x N-AnkR C-TRP box-C-P olyP (EWKFAR…PP P FN) C-CIRB

3x N-AnkRs C-TRP box-C-P olyP (EWKFAR…PP P FN) C-CIRB

Reporte d Interac ting Prote ins

TRPC4 TRPC5 (TRP C3 -embr yo), Ca lmodulin, PC2 (TRPP1) IP3Rs, cave olin-1, Ga q/11, PMCA

TRPC 1(e mbryo only) PLCb, P LCg, P3R, RyR, SERCA, cav e olin-1, Calmodul in aSNAP, NC X1

TRPC1 , TRP C5 PLCb; NHERF-2, NHERF-1 an d ZO-1 via PDZ-BD EBP50( ez rin/moes in/radixin-binding p hos phoprotein 50) IP3R, calmo dulin (TRP C4a ), TRPC 4a ac ts as dominant ne gative for TRPC 4b

TRPC1 , TRP C4 calmodul in

TRPC3 , TRP C7, calmodul in, FKBP12

TRPC3 , TRP C6, calmodul in, FKBP12

Activa tion

Mus car inic M1 GPCR, Gq-PLCb TKR-PLCg?

GPCR Gq-PLCb, DAG, TKR-PLCg?

Hista mine H1 GPCR Gq-P LCb;DAG TKR-PLCg [Ca 2+]I?

Mus car inic M1 GPCR Gq-PLCb TKR-PLCg? 100 µM La3+ Calmida zolium

Mus car inic M1GP CR;Gq-PLCb TKR-PLCg? 100 µM La3+

Hista mine H1 GPCR Gq-PLCb,DAG TKR-PLCg?, [Ca 2+]I via CaMKII, AlF4, 100 µM flufe na mate, 100 µM niflumate, 100 µM GTP S

HEK pur ine rgic GPCR Gq-PLCb, DAG, TKR-PLCg? [Ca 2+]I 100 µM GTP S

Direct Agonist

N.D. N.D. N.D. N.D. N.D. N.D.

~P Ca/PNa <10 N.D. 1.6 ~1

~1

5 ~0.5

Blo c kers and inhibitors

80 µM 2-APB, 10 µM Gd, La Ca 2+-CaM inhibits

N.D. 2-APB, 1-25 µM Gd, La, RuR

mM La3+ ; 2-APB Niflumic ac id, DIDS Increa s ed [Ca 2+]I (Walke r)

mM La3+ ; 2-APB (IC50) 0. 25 mM Cd, 4 µM La , 2 µM Gd, 130 µM amiloride, 4 µM SKF9 6365, 100 µM 2 -APB bloc ks ~60% of current, Ca e ma y block as perme ate s .

100 µM La 3+ SKF9 6365 (25 µM), 0.1-0.4 mM Ca e , Ca i via ca lmodulin

Sin g le channel g

10 pS in 1 nM Ca 2.5 pS in 2 [Ca 2+]I Va ca JBC 2002 ~30 pS a t –50 mV a nd ~5 pS at 50 mV (s ymmetrical 200 mM K+ with 40 mM internal Mg) Maroto et al, NCB, 2005

42pS (symmetr ical 140 mM Na) Luca s e t al, Ne uron 2003

Uncle ar as homomer. Estimates r a nge from ~20 pS (nois e anal ysis) to 65 pS (140 Na /140 Na)?

Uncle ar as homomer- Estimates r a nge from 18 pS to 40 pS 18 pS (b) Walke r, AJP) 28 pS (Scha e fer) 140 Na/140Cs)

~38 pS as homome r ~5 pS as TRPC1 /TRP C5 he terome r

35 (sym 120 Cs , Hofma nn, 99 ); 28 pS isotonic (Mori)

Rectifying o pen cha nne l I-V: 50 pS (0 to +100 mV), 25 pS (-100 to 0 mV) (Shi, 2004)

I-V s hape

Nea r line ar

From Lucas et a l, Neu ron 2 003

From Dietr ich et a l. JBC

~ Double -re ctifying as hom omer

From Scha efe r e t al. JBC

~ Double -re ctifying as hom omer

from Jung et al, AJP

Shi et al, J Ph ys iol 2004

Comm e nts Mos t un ique of TRPC seq uences

No funct ion in huma n. TRPC2 b is maj or varia nt on mous e cell memb rane (VNO)

Ge nerally a cce pte d role for DAG activa tion. “S tore ope rat ion” in dispute.

TRPC4 -/- mou s e a orta en dothelial cells h a d red uce d “CRAC-like” “store -ope ra ted ” current, le ad ing to an impaire d en dothelium-de pe nde nt rela xation of blood vessel s . 5-HT med iate d rel e as e of GABA from thala mic inte rneu rons is gre atly re duced in TRPC4 -/- mice . Ove rall mous e phe notype not reported . Majority of eviden ce indica tes that TRP C4b is recep tor-ope rated .

G p rotein linked rec e ptor o pe ra ted cha nne l as he teromer . I/V of he teromer is simila r to NMDA clas s of ion channels (ne gat ive s lope region), but unlike NMDA cha nnels is not de pende nt on Mg volta ge -de pe ndent rectificat ion. Not st ore -ope rated .

Ge nerally a cce pte d role for DAG activation. Pro bable a1- ad renoce ptor a ctiva ted cha nne l in po rtal vein smooth muscle , as well as o the r vascu lar smoo th musc le. PLC act ivat ion, OAG a nd tha psigarg in st imulate delive ry of cha nne ls to plas ma me mbra ne

Ge nerally a cce pte d role for DAG activa tion

Maroto et al Nat Cell Biol 2005

The TRPM Melastatin Subfamily TRP Subtype Unigene

M1 (MLSN) Hs.43265 Alt spliced

M2 (hTRPC7, LTRPC2) Mm.133517

M3 (hKIAA1616, LTRPC3) Hs.288911

M4 (hFLJ20041, LTRPC4, CAN 4b]) Hs.31608

M5 (Mtr1, LTRPC5) Hs.272287

M6 (CHAK2) Hs.272225

M7 (CHAK1, TRP-PLIK, LTRPC7) Hs.33819

M8 (Trp-p8, CMR1) Hs.366053

# Amino acids

1533 1503 (splice variants) 1325 (Lee et al) 1555 (Grimm et al) (up to 6 splice variants)

1214 (M4b) 1040 (M4a) 678 (M4c)

1165 2022 1864 1104

OMIM gene map (h) (m)

15q13-q14 21q22.3 9q21.11 19q13.33 11p15.5 9q21.13 15q21 2q37.1

Genetic model or disease mutation

Mouse TRPM5-/-

Human hypomagnesemia with secondary hypocalcemia (HGH)

Known Functions

Taste (Sweet, Bitter, Umami)

Renal and gastrointestinal tract Mg2+ uptake

Noxious cold

Likely role in

Melanoma invasiveness

Oxidant stress response in immune and other cells (microglia in brain, respiratory burst in neutrophils?)

Ca2+ absorption in renal collecting tubule

Regulation of calcium oscillations in activated T lymphocytes Myogenic cerebral artery constriction

Magnesium absorption in kidney

Cellular Mg homeostasis proposed. Potential other trace metals, such as Zn, Mn. Oxygen-glucose deprivation-induced calcium influx and neuronal death

Pain pathways (present in A fibers), presynaptic potentiation in DRG neurons

N or C terminal domain modules

N-700 a.a. TRPM domain, W/FIX3-(F/l/I)XK(R/K)EC(V/I/S)X12-

24CXCG C-TRP (VWKFQR) box C-PolyP C-coiled coil

N-700 a.a. TRPM domain, W/FIX3-(F/l/I)XK(R/K)EC(V/I/S)X12-24CXCG [VIYFL]WK[FAY][QN]R; C-NUDT9 ADP-ribose hydrolase (NUDt9-H) C-TRP (IWKFQR) box C-PolyP;C-coiled coil

N-700 a.a. TRPM domain, W/FIX3-(F/l/I)XK(R/K)EC(V/I/S)X12-24CXCG C-TRP (VWKFQR) box [VIYFL]WK[FAY][QN]R; C-PolyP C-coiled coil

N-700 a.a. TRPM domain, W/FIX3-(F/l/I)XK(R/K)EC(V/I/S)X12-24CXCG C-TRP (YWKAQR) box [VIYFL]WK[FAY][QN]R; C-PolyPi

C-coiled coil

C-TRP (FWKFQR) box [VIYFL]WK[FAY][QN]R; C-PolyP C-coiled coil

N-700 a.a. TRPM domain W/FIX3-(F/l/I)XK(R/K)EC(V/I/S)X1

2-24CXCG C-TRP (LWKYNR) box [VIYFL]WK[FAY][QN]R; C-PolyP C-coiled coil C-Atypical protein kinase

N-700 a.a. TRPM domain, W/FIX3-(F/l/I)XK(R/K)EC(V/I/S)X12-

24CXCG C-TRP (VWKYQR) box [VIYFL]WK[FAY][QN]R; C-PolyP C-coiled coil

N-700 a.a. TRPM domain, W/FIX3-(F/l/I)XK(R/K)EC(V/I/S)X12-

24CXCGC-TRP (VWKFQR) box [VIYFL]WK[FAY][QN]R; C-PolyP C-coiled coil

Reported Interacting Proteins

Short transcript of M1 (MLSN-S).

N.D. N.D. N.D. N.D. TRPM7 PLC PLC 1 TRPM6, annexin 1

Activation N.D. ~100 µM ADP Ribose mM NAD, NAD mM H2O2

Constitutive, Enhanced by hypoosmolarity Activated by sphingolipids

M4b:Kact: 300-500 nM [Ca2+]I; >1 µM (Nilius)

GPCR (T1R, T2R) Ggus-PLC 2 [Ca2+]I 1-100 µM

PIP2

PIP2 hydrolysis by GPCR (M1, LPAR/EDG-7R)-Gq-PLC and TK(EGF) -PLC hydrolyzes PIP2 to turn off channel. Tyrosine phosphorylation, Activation upon oxidative stress and cAmp proposed, PKA

T<25oC (80 µM menthol, icilin potentiate)

Direct Agonist (bind channel protein to activate)

N.D. Probably ADP Ribose Menthol?

~PCa/PNa ~1 (estimated) 1.6 M4b: <0.05 (monovalent selective)

<0.05 (Monovalent selective)

<1 0.3 monovalents outward inward: Zn~Ni>Ba>Co>Mg~Mn~Sr~Cd~Ca

1-3

Blockers and inhibitors

N.D. N.D. 100 µM Gd M4a: 80 µM La, Gd M4b: intracellular free ATP 4-, 1.7µM

0.6 mM [Mg2+]i

Single channel

. .N D 60 (140 /140 )pS Na Na 83 (140 /140 )Na Na 4 :25 (M b pS isotonic )monovalents

23 (pS isotonic monovalents)S

. .N D 105 (+ )pS voltages

83 (+ )pS voltages

I - V shape . .N D

/ Instantaneous I V is linear

From Perraud et .al Nature

FromGrimm et .al JBC

4 :M b

/ Instantaneous I V is

, , 2003linear Nilius et al

Hofmann

. From Voets et al JBC

Runnels et al NCB

From Mckemy et al. Nature

Comments Decrease in M1 RNA with melanoma progression

Increased [Ca2+] I potentiates (EC50 340 nM)

Independent of [Ca2+] I Insensitive to 1 µM RuR

Probably ubiquitous Voltage-modulated calcium-activated (VMCA) is an unusual feature. Decavanadate (2µM) inhibits voltage dependence

Candidate gene for rhabdomyosarcomas, Wilms tumor, Beckwith-Widemann Voltage-modulated Calcium-activated (VMCA) is an unusual feature

Contains a protein kinase on its C terminus as part of the protein (Chanzyme) Currently difficult to separate from TRPM7 current measurements

Contains a protein kinase on its C terminus as part of the protein (Chanzyme). The kinase activity is not required for channel conduction. Overexpression results in cell death. Deletion also results in cell death in DT40 cells. Kinase domain phosphorylates Annexin 1

Correlated with trkA NGF receptors trp-p8 mRNA also expressed in primary tumors of prostate, breast, colon, lung, and skin. Loss of prostate trp-p8 mRNA e associated with shorter survival. trp-p8 may be androgen- regulated.

TRPM Family

The TRPV Vanilloid Receptor Subfamily TRP Subtype Unigene

V1 (VR1) Hs.283010

V2 (VRL1,GRC,OTRPC2) Hs.279746

V3 Hs.375034

V4 (OTRPC4, VR-OAC) Hs.287776

V5 (ECaC1, CaT2) Hs.283369

V6 (ECaC2, CaT1) Hs.302740

# Amino acids 839 764 791 871 729 725 OMIM gene map (h) 17p13.3 17p11.2 17p13.3 12q24.1 7q3S 7q33-34 Genetic model or disease mutation

Mouse TRPV1-/- Mouse TRPV4-/-

Known Functions

Basis of “hot” pepper sensation Heat evoked nociception Inflammatory thermal hyperalgesia, Bladder distension sensing Chronic cough, bronch ial sensiti zat ion

Thermal pain (medium diameter neurons); Trigeminal neuron, pterygopalatine and submandibular ganglia, myenteric plexus & nodose ganglion neurons, spinal cord lamina I, II, hypothalamic paraventricular, suprachiasmatic, and supraoptic nuclei, retina Osmo-sensing in aortic myocytes

CNS osmotic sensing (circumventricular) Pressure sensing (DRG) Nociception in primary afferents sensitized by PGE2 Inflammatory thermal hyperalgesia,

Mouse TRPV5-/-

Likely role in Other pain pathways Skeletal and cardiac muscle degeneration Other pain pathways (visceral sensory) Present in cranial nerve ganglia, spinal cord, murine aortic myocytes

Warm temp sensing in 308 ketatinocytes Pain

Temperature sensing in skin, 308 ketatinocytes

Ca2+ uptake in kidney (Vit D regulated)

Ca2+ uptake in intestine (Vit D regulated)

N or C terminal domain modules

3x N-AnkR C-TRP (IWKLQR) box C- Putative PIP2 binding site

3x N-AnkR C-TRP (IWKLQK) box

3x N-AnkR C-TRP (IWRLQR) box

4x N-AnkR C-TRP (IWKLQW) box

5x N-AnkR C-TRP (LWRAQV) box N-PDZ

4x N-AnkR C-TRP (LWRAQI) box N-PDZ

Reported Interacting Proteins

TRPV3 Calmodulin Snapin, synaptotagmin IX

-tubulin

RGA protein TRPV1 Calmodulin (C term) Src family kinases (potentiation)

TRPV6 S100A10/annexin 2

TRPV5 S100A10/annexin 2

Activation T>43oC Capsaicin,(0.7 µM) Resiniferatoxin; 40 nM Anandamide, ACEA satie ty fa ctor oleo ylethanola mide H+, 2APB ~200µM strongly modulated by voltage.

T>53oC IGF-1 2-APB ~100 M Neuropeptide head activator

T>30oC 2-APB 30µM

Decreased Osmolarity (260-200 mOsm) T>24oC Phorbol esters

Constituitive [Ca2+]I inhibits

Constituitive [Ca2+]I inhibits

Q10 ~20 at negative voltages, Q10 is much smaller at positive voltages

N.D. 25 10,19 N.D. N.D.

Direct Agonist (bind channel protein to activate)

0.7 µMCapsaicin 40 nMResiniferatoxin Anandamide, H+ Eicosanoids?

Capsaicin Insensitive Capsaicin Insensitive

Capsaicin Insensitive Endocannabinoids (anandamide), arachidonic acid, epoxyeicosatrienoic acids

~PCa/PNa 10 (5 for PMg/PNa) 3 ~10 6 >100 (Ca>Sr~Ba>Mn)

>100 (Ca>Ba>Sr>Mn)

Blockers and inhibitors

Capsazepine 10 µM RuR; PIP2

1 µM RuR, La3+, SKF96365

100 nM RuR La3+

100 nM RuR, 100 µM Gd3+, 0.5 mM La3+

120 nM RuR µM Cd>>Gd>La, Mg Cu, Pb

9 µM RuR

µM Cd>>Gd>La Mg

Single channel ~80 (+ , 140 pS V/140 )Na Na

. . N D 170 ,pS conductance is

dependent ontemperature with

low Q10

90 (+60 , pS mV140 /140 )Na K

75 (pS Ca-f )ree 42-58 (pS Ca-f )ree

I - V shape

Julius

From Caterina e t al. Nature

Xu et al

FromWatanabl e et al. JBC

-100 100-300-200-100 pAmV

Courtsey to Kirichok

Yue e t al, Nature 2001

Comments PKC enhances activation

PKA regdesensitization

Acid potentiates Heat sensitizes

Not store-activated Activation ofP LC

releases V1 from PIP2 inhibition

Capsaicin induced neuronal death

Difficult .expression Most . data from oocytes IGF-1

2appears to translocate TRPV .to membranes in mouse

2 Heat sensitizes TRPV on repetitive stimulation

Current displays sensitization with

repeatedtemperature

ramps and2 APB.activation

Marked hysteresis of

temperature (reponse gradual , increase with T

sharp decline with cooling)

Most Ca2+- selective of TRPs- conserved asp in

.pore Voltage-

[dependent Mg2+]i-block

Most Ca2+- selective of TRPs-

conserve d Asp in.pore

Voltage- [dependent Mg2+]i-

block

TRPV Family

Distant TRPs

Distant TRPs (A,P,ML) TRP Subtype Unigene

A1 (ANKTM1) Mm. 186329

P1 (PC2, PKD2) Hs.82001

P2 (PKD2L1) Hs. 159241

P3 (PKD2L2) Hs.272418

ML1 (MCOLN1) Hs.12909

ML2 (MCOLN2) Hs.210792

ML3 (MCOLN3) Hs.49344

# Amino acids 1126 968 805 609 580 566 553

OMIM g ene map (h) (m)

8q13 4q21-q23 10q24 5q31 19p13.2-13.3 1p22.3 1p22.3

Genetic model or disease mutation

N.D. Polycystic kidney Disease (ADPKD) Mouse Pkd2-/-

krd mice (P2-/-) N.D. Defective in mucolipidosis type IV, lysosomal storage disease

Varitint-waddler mouse (Va) A419P in 5th TM domain

Known Functions

Mouse cardiac, skeletal and renal development,; kidney& liver cyst formation With PC2(PKD2) mechanosensation in cilia (kidney);

Kidney development (mice)

N.D. Cell vacuolar defects- probably defect in sorting/transport in late endocytic pathway

Hair cell , stereocilia maturation

Likely role in Sensory in trigeminal and DRG, Putative “hearing” channel in hair cells of cochlea

intracellular channel, fertil ity

retinal development (mice)

Endosomes Intracellular vesicle defect

N or C terminal domain modules

14 N-AnkR C-EF hand + C-coiled coil

N-proline Leucine zipper S2 Nuclear local. aa 43-60 C-di-leucine

Reported Interacting Proteins

N.D. PC1(PKD1), Hax-1, Cortactin Cadherin via PC1

Activation Mustard oils, PLC-coupled GPCRs, Icilin, cannabinoids T<18oC?

Mechanical stress? [Ca2+] i?

~1 µM [Ca2+] i [Ca2+] i ? [Ca2+] I? N.D. N.D.

Direct Agonist 100 µM Icilin ~PCa/PNa 1.4

PCa~PMg ~1-5 ~4 ~1-5,

PMg~0 ~1

Blockers and inhibitors

5 µM RuR

Amiloride (30 µM) Gd, La

Amiloride (130 µM) mM Gd, La Ni

Single channel

. . N D ~40 pS highCao( ), ~50 Luo et al

140pS KGonzalez

137 (100 pS mM) Na Chen et al

300 ?pS 83 (100 ) mM NaLaPlante

I - V shape

2003Story et al

Polyc 1ystin+ 2; P (Linear Germino et

2000)al

~linear

. From Chen et alNature

. .N D . . N D . . N D . . N D

Comments Sensitive to plant-derived irritants such as mustard ,oils

and perhaps to/membrane cytoskeletal

perturbations such as stretch (as the hearing transduction

). channel May depolarize nociceptors in response to pro-

( .algesic agents e g. ,bradykinin , ) serotonin histamine that

activate PLC-coupled recepto .rs Originally proposed to sense painfully cold temperatu .res

Decreased in fibroblasts afteroncogen ic transformation

1In P -/- , cell delayed cargo exit

from the Golgi apparatus

Role in/sortingtransport in

late endocytic, pathway defective

in mucolipidosis type IV lysosomal

storage disease

Mouse- Early- onset ,hearing loss

, vestibular defectspigmentation abnormalities