Crystal structure of human stem cell factor: Implication for stem cell factor receptor dimerization and activation

Zhang, Zhongtao; Zhang, Rongguang; Joachimiak, Andrzej; Schlessinger, Joseph; Kong, Xiang‐Peng

doi:10.1073/pnas.97.14.7732

Cited by 119 publications

(104 citation statements)

References 49 publications

(49 reference statements)

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“…Structural analyses of SCF dimers have shown that complex formation between the two SCF protomers is mediated by a large interface involving primarily hydrophobic interactions (8,9). Moreover, on KIT binding, the angle between the two SCF protomers is altered by ∼5° (10, 11).…”

Section: Resultsmentioning

confidence: 99%

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Structure, domain organization, and different conformational states of stem cell factor-induced intact KIT dimers

Opatowsky

Lax

Tomé

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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Using electron microscopy and fitting of crystal structures, we present the 3D reconstruction of ligand-induced dimers of intact receptor tyrosine kinase, KIT. We observe that KIT protomers form close contacts throughout the entire structure of ligand-bound receptor dimers, and that the dimeric receptors adopt multiple, defined conformational states. Interestingly, the homotypic interactions in the membrane proximal Ig-like domain of the extracellular region differ from those observed in the crystal structure of the unconstrained extracellular regions. We observe two prevalent conformations in which the tyrosine kinase domains interact asymmetrically. The asymmetric arrangement of the cytoplasmic regions may represent snapshots of molecular interactions occurring during trans autophosphorylation. Moreover, the asymmetric arrangements may facilitate specific intermolecular interactions necessary for trans phosphorylation of different KIT autophosphorylation sites that are required for stimulation of kinase activity and recruitment of signaling proteins by activated KIT.receptor tyrosine kinases | cell signaling | cancer | structure analysis | structural biology A broad range of fundamental cellular processes, including proliferation, differentiation, survival, and metabolism, are mediated by signaling pathways that are activated by the 58 members of the receptor tyrosine kinase (RTK) family of membrane receptors (1). The receptor tyrosine kinase KIT was initially discovered as the viral oncogenic protein of a feline sarcoma virus (2). Subsequent studies have demonstrated that KIT and its ligand stem cell factor (SCF) play important roles in the control of proliferation, differentiation, and survival of a variety of cell types, including germ cells, hematopoietic cells, melanocytes, intestinal pacemaker cells, and sensory neurons in the CNS (3-5).KIT and other members of type-III RTK family are composed of an extracellular ligand-binding region containing five Ig-like domains (designated D1-D5), followed by a single transmembrane (TM) spanning helix, a relatively large (35 aa) cytoplasmic juxtamembrane (JM) region, and a tyrosine kinase domain with a kinase insert region containing several autophosphorylation sites. Structural and biochemical studies have shown that the N-terminal region composed of D1, D2, and D3 of KIT extracellular region functions as a binding site for SCF, which, by virtue of its homodimeric structure, is responsible for bringing about KIT dimerization (6-11).SCF-induced KIT dimerization results in homotypic contacts between the membrane proximal Ig-like domains D4 and D5 of two neighboring KIT molecules. These homotypic contacts position the TM and the cytoplasmic regions of KIT and other RTKs at a distance and orientation that facilitate tyrosine autophosphorylation, stimulation of enzymatic activity, and cell signaling (11-13). Oncogenic gain-of-function mutations in KIT have been identified in various cancers, including gastrointestinal stromal tumors, acute myeloid leukemia, melanoma, an...

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Section: Resultsmentioning

confidence: 99%

“…Structural and biochemical studies have shown that the N-terminal region composed of D1, D2, and D3 of KIT extracellular region functions as a binding site for SCF, which, by virtue of its homodimeric structure, is responsible for bringing about KIT dimerization (6)(7)(8)(9)(10)(11).…”

mentioning

confidence: 99%

Structure, domain organization, and different conformational states of stem cell factor-induced intact KIT dimers

Opatowsky

Lax

Tomé

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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“…The soluble protein is heavily N-and O-glycosylated, has extensive secondary structures and is dimeric (non covalently associated) under non-denaturating conditions (Arakawa et al, 1991;Langley et al, 1992;Zhang et al, 2000). The presence or absence of the carbohydrate moieties does not influence the biological activity (Flanagan and Leder, 1990;Zsebo et al, 1990), whereas the two intramolecular disulfide bonds of SCF are essential (Jones et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

Expression and purification of bioactive soluble murine stem cell factor from recombinant Escherichia coli using thioredoxin as fusion partner

Bals

Schambach

Meyer

et al. 2011

Journal of Biotechnology

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“…These processes occur through activation of KIT upon binding by stem cell factor (SCF), a ligand found in membrane-anchored and soluble forms (1,2) in a variety of cell types, including hematopoietic stem cells, germ cells, vascular endothelial cells, and the mesenchymal cells with uniquely neuromuscular differentiation known as the interstitial cells of Cajal (3,4). KIT belongs to the type III subfamily of RTKs (5), a family composed of an extracellular region that includes five Ig-like domains (designated D1-D5), a single transmembrane domain (TM), a juxtamembrane region (JM), a tyrosine kinase domain split by a kinase insert, and a C-terminal tail (6) (Fig.…”

mentioning

confidence: 99%

Structural basis for KIT receptor tyrosine kinase inhibition by antibodies targeting the D4 membrane-proximal region

Reshetnyak

Nelson

Shi

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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Somatic oncogenic mutations in the receptor tyrosine kinase KIT function as major drivers of gastrointestinal stromal tumors and a subset of acute myeloid leukemia, melanoma, and other cancers. Although treatment of these cancers with tyrosine kinase inhibitors shows dramatic responses and durable disease control, drug resistance followed by clinical progression of disease eventually occurs in virtually all patients. In this report, we describe inhibitory KIT antibodies that bind to the membrane-proximal Ig-like D4 of KIT with significant overlap with an epitope in D4 that mediates homotypic interactions essential for KIT activation. Crystal structures of the anti-KIT antibody in complex with KIT D4 and D5 allowed design of affinity-matured libraries that were used to isolate variants with increased affinity and efficacy. Isolated antibodies showed KIT inhibition together with suppression of cell proliferation driven by ligand-stimulated WT or constitutively activated oncogenic KIT mutant. These antibodies represent a unique therapeutic approach and a step toward the development of "naked" or toxin-conjugated KIT antibodies for the treatment of KIT-driven cancers.phosphorylation | therapeutic antibodies | cancer therapy | cell signaling | protein kinase

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Crystal structure of human stem cell factor: Implication for stem cell factor receptor dimerization and activation

Cited by 119 publications

References 49 publications

Structure, domain organization, and different conformational states of stem cell factor-induced intact KIT dimers

Structure, domain organization, and different conformational states of stem cell factor-induced intact KIT dimers

Expression and purification of bioactive soluble murine stem cell factor from recombinant Escherichia coli using thioredoxin as fusion partner

Structural basis for KIT receptor tyrosine kinase inhibition by antibodies targeting the D4 membrane-proximal region

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