Potent human p140-TrkA agonists derived from an anti-receptor monoclonal antibody

LeSauteur, Lynne; Maliartchouk, Sergei; Jeune, H Le; Quirion, Rémi; Saragovi, H. Uri

doi:10.1523/jneurosci.16-04-01308.1996

Cited by 63 publications

(74 citation statements)

References 37 publications

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“…Primary Antibodies-Anti-TrkA mouse mAb 5C3 binds to the TRKA-D5 subdomain with K d ϳ 2 nM and acts as a full agonist of this receptor (22). Mouse NGF30 mAb is an anti-NGF mAb that when bound to NGF still allows high affinity binding of NGF to TrkA.…”

Section: Methodsmentioning

confidence: 99%

“…Therefore, prior to binding studies, we used mAb 5C3 (22) that binds a conformationally sensitive epitope near the NGF-binding site of these recombinant proteins. ELISA studies were done to quantify appropriately folded TrkA-ECD and TRKA-D4 -D5 as described previously (23).…”

Section: Methodsmentioning

confidence: 99%

“…The binding of anti-TrkA mAb 5C3, an agonistic TrkA ligand that binds at the D5 subdomain and saturates TrkA at 65 nM (22), was blocked by NGF and NT-3 to different levels. NGF at 50 nM inhibited ϳ20% of 5C3 binding (Fig.…”

Section: Chimeras Exhibit Trophic Signals In Response Tomentioning

confidence: 99%

“…I-NGF Binding Studies on HEK293 Cells Expressing 4.1 Chimeric Receptors-Scatchard plot analysis of 125 I-NGF binding to cells was done as described (22,23). Viable HEK293 cells expressing the 4.1 chimera (with TrkB-D5 subdomain replaced by subdomain D5 of TrkA) were used as test, and viable wild type HEK293 cells were used to determine nonspecific 125 I-NGF binding.…”

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confidence: 99%

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TrkA Receptor “Hot Spots” for Binding of NT-3 as a Heterologous Ligand

Ivanisevic

Zheng

Woo

et al. 2007

Journal of Biological Chemistry

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Neurotrophins signal via Trk tyrosine kinase receptors. Nerve growth factor (NGF) is the cognate ligand for TrkA, the brainderived neurotrophic factor for TrkB, and NT-3 for TrkC. NT-3 also binds TrkA as a lower affinity heterologous ligand. Because neurotrophin-3 (NT-3) interactions with TrkA are biologically relevant, we aimed to define the TrkA "hot spot" functional docking sites of NT-3. The Trk extracellular domain consists of two cysteine-rich subdomains (D1 and D3), flanking a leucinerich subdomain (D2), and two immunoglobulin-like subdomains IgC1(D4) and IgC2(D5). Previously, the D5 subdomain was defined as the primary ligand-binding site of neurotrophins for their cognate receptors (e.g. NGF binds and activates through TRKA-D5 hot spots). Here binding studies with truncated and chimeric extracellular subdomains show that TRKA-D5 also includes an NT-3 docking and activation hot spot (site 1), and competition studies show that the NGF and NT-3 hot spots on TRKA-D5 are distinct but partially overlapping. In addition, ligand binding studies provide evidence for an NT-3-binding/allosteric site on TRKA-D4 (site 2). NT-3 docking on sites 1 and/or 2 partially blocks NGF binding. Functional survival studies showed that sites 1 and 2 regulate TrkA activation. NT-3 docking on both sites 1 and 2 affords full agonism, which can be additive with NGF activation of Trk. However, NT-3 docking solely on site 1 is partially agonistic but noncompetitively antagonizes NGF binding and activation of Trk. This study demonstrates that Trk signaling is more complex than previously thought because it involves several receptor subdomains and hot spots.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Chimeras Exhibit Trophic Signals In Response Tomentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

TrkA Receptor “Hot Spots” for Binding of NT-3 as a Heterologous Ligand

Ivanisevic

Zheng

Woo

et al. 2007

Journal of Biological Chemistry

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“…We have previously described a monoclonal antibody (mAb) 5C3 that binds a restricted epitope of TrkA with high affinity and acts as a full agonist (when compared with NGF) on cells that express TrkA but do not express p75 (LeSauteur et al, 1996). In the present study, combinations of the TrkA-specific mAb 5C3 and the p75-specific mAb MC192 (Chandler et al, 1984) were used as ligands to analyze NGF receptor in functional and biochemical assays.…”

Section: ϫ10mentioning

confidence: 97%

Optimal Nerve Growth Factor Trophic Signals Mediated by Synergy of TrkA and p75 Receptor-Specific Ligands

Maliartchouk¹,

Saragovi

1997

J. Neurosci.

107

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Growth factor receptors: Structure, mechanism, and drug discovery

McInnes

Sykes

1997

Biopolymers

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The focus of this review is the relationship between the three‐dimensional structure of ligands of the various members of the growth factor receptor tyrosine kinase superfamily and their interaction with the cognate receptor. Particular attention is given to the transforming growth factor—α, epidermal growth factor (EGF); nerve growth factor, neurotrophin; and insulin‐like growth factor—1 (IGF‐1), insulin systems since these have been extensively studied in recent years. The three receptor types, which bind these ligands, are the epidermal growth factor receptor family (erb B receptors), the neurotrophin or Trk receptor family, and IGF‐1/insulin receptors, respectively, and represent three distinct members of the tyrosine kinase superfamily. For each of these, formation of the ligand‐receptor complex initiates the signal transduction cascade through autophosphorylation by the intracellular tyrosine kinase domain. The extracellular portion of the receptor that contains the ligand binding domain in these systems varies significantly in organization in each case. For the EGF receptor system, ligand binding induces homo‐ and heterodimerization of the receptor leading to activation of the intracellular kinase. For the Trk receptor system, homodimerization of receptors has been shown to occur, although a second receptor, p75, is also required for high affinity binding of neurotrophins and for enhanced sensitivity of tyrosine kinase activation at low ligand concentrations. The IGF‐1 and insulin receptors exist as covalent cross‐linked dimers where each monomer is composed of two subunits. The aim of this review is also to discuss the mechanism of ligand‐receptor interaction for each of these cases; however, since no structural information is yet available for the ligand‐receptor complex, the discussion will largely be centered on the molecular requirements of ligand binding. As these receptors are activated through the ligand binding site on the extracellular domain, this represents a possible target for pharmacological intervention by inhibition or stimulation of this portion of the receptor. Thus from a drug design perspective, the focus of this review is to discuss progress in the development of agonists or antagonists of the ligand for these receptors. © 1998 John Wiley & Sons, Inc. Biopoly 43: 339–366, 1997

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Potent human p140-TrkA agonists derived from an anti-receptor monoclonal antibody

Cited by 63 publications

References 37 publications

TrkA Receptor “Hot Spots” for Binding of NT-3 as a Heterologous Ligand

TrkA Receptor “Hot Spots” for Binding of NT-3 as a Heterologous Ligand

Optimal Nerve Growth Factor Trophic Signals Mediated by Synergy of TrkA and p75 Receptor-Specific Ligands

Growth factor receptors: Structure, mechanism, and drug discovery

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