Amy Stiegler, PhD
Senior Research Scientist in PharmacologyCards
About
Research
Publications
2025
Nickel Binding to the c-Src SH3 Domain Facilitates Crystallization
Calicdan X, Fisher O, Ha B, Boggon T, Stiegler A. Nickel Binding to the c-Src SH3 Domain Facilitates Crystallization. Protein And Peptide Letters 2025, 32: 679-692. PMID: 41088944, PMCID: PMC12582252, DOI: 10.2174/0109298665417324250929120040.Peer-Reviewed Original ResearchC-Src SH3 domainSH3 domainC-SrcCrystal structureCrystallization bufferPseudo-twofold rotation axisX-ray crystal structureSpace group H32Nickel bindingSrc SH3 domainAmino acid sequenceCrystal formsUnique crystal structureSurface-exposed histidine residuesOrdered water moleculesCrystal lattice contactsMultiple crystal formsAmino-terminal copperMetal-mediated crystallizationCrystal packingAtomic-level informationAcid sequenceWater moleculesNickel coordinationLattice contactsSHP2 genetic variants in NSML-associated RASopathies disrupt the PZR–IRX transcription factor signaling axis
Perla S, Stiegler A, Yi J, Enyenihi L, Zhang L, Riaz M, An E, Qyang Y, Boggon T, Bennett A. SHP2 genetic variants in NSML-associated RASopathies disrupt the PZR–IRX transcription factor signaling axis. Proceedings Of The National Academy Of Sciences Of The United States Of America 2025, 122: e2503631122. PMID: 40854126, PMCID: PMC12415285, DOI: 10.1073/pnas.2503631122.Peer-Reviewed Original ResearchConceptsKnock-in mutationSH2 domainGenetic variantsSH2 domains of SHP2Iroquois homeoboxBinding to SHP2NSML miceProtein tyrosine phosphataseHypertrophic cardiomyopathyExpression levelsPhosphorylation-deficientProtein expressionCellular SrcTranscription factor 3Tyrosine phosphataseSHP2Protein zeroRare autosomal dominant disorderLow-dose dasatinibAutosomal dominant disorderPostnatal cardiac growthExpression of Irx3Signaling AxisIRX3Factor 3Complement factor I deficiency–associated neuroinflammatory disease among Old Order Amish
Reid W, Baas L, Stiegler A, Boggon T, Kreiger P, Sullivan K, Behrens E, Kandula V, van den Heuvel L, Brigatti K, Carson V, Romberg N. Complement factor I deficiency–associated neuroinflammatory disease among Old Order Amish. The Journal Of Allergy And Clinical Immunology 2025, 156: 835-841. PMID: 40602690, DOI: 10.1016/j.jaci.2025.06.021.Peer-Reviewed Original ResearchConceptsCerebral spinal fluid pleocytosisSpinal fluid pleocytosisInborn disorderClinical response to eculizumabSpine magnetic resonance imagingResponse to eculizumabSingle-center cohortAcute disseminated encephalomyelitisOld Order Amish populationMagnetic resonance imagingTransverse myelitisDisseminated encephalomyelitisFemale predominanceHomozygous patientsLeukocyte frequenciesLive birthsAseptic meningoencephalitisCytokine concentrationsDiagnostic considerationsAmish populationComplement consumptionPatient samplesClinical laboratoriesComplement regulatorsNeuroinflammatory diseasesNovel heterozygous SPI1c.538C>T p.(Leu180Phe) variant causes PU.1 haploinsufficiency leading to agammaglobulinemia
Daddali R, Kettunen K, Turunen T, Knox A, Laine P, Chowdhury I, Vänttinen M, Mamia N, Stiegler A, Boggon T, Kere J, Romberg N, Seppänen M, Varjosalo M, Martelius T, Grönholm J. Novel heterozygous SPI1c.538C>T p.(Leu180Phe) variant causes PU.1 haploinsufficiency leading to agammaglobulinemia. Clinical Immunology 2025, 277: 110503. PMID: 40294836, DOI: 10.1016/j.clim.2025.110503.Peer-Reviewed Original ResearchConceptsDendritic cell countHeterozygous loss-of-function variantsLoss-of-function variantsCentrosome-associated proteinsFamily transcription factorsEts family transcription factorGene expression patternsAssociated with B cell developmentHematopoietic cell fatePDC countsB cell developmentCell countDisrupts gene expression patternsB-cell countsCell fateCarrier sisterSevere bacterial infectionsTranscription factorsIsolated IgA deficiencyFinnish familiesExpression patternsIn vitro studiesIgA deficiencyVariable penetranceIndex patientCancer hotspot mutations rewire ERK2 specificity by selective exclusion of docking interactions
Robles J, Stiegler A, Boggon T, Turk B. Cancer hotspot mutations rewire ERK2 specificity by selective exclusion of docking interactions. Journal Of Biological Chemistry 2025, 301: 108348. PMID: 40015635, PMCID: PMC11982978, DOI: 10.1016/j.jbc.2025.108348.Peer-Reviewed Original ResearchShort linear motifsCancer hotspot mutationsLinear motifsERK substratesYeast two-hybrid libraryHotspot mutationsTwo-hybrid libraryCancer-associated mutantsDocking interactionsWild-type ERK2Cancer-associated mutationsDocking motifBinding sequenceKinase ERK2Co-crystal structureMutant formsERK2 mutantsDisordered regionsERK2MotifStructural rationalePeptide bindingMutationsWT kinasePeptide fragmentsThe C2 domain augments Ras GTPase-activating protein catalytic activity
Paul M, Chen D, Vish K, Lartey N, Hughes E, Freeman Z, Saunders T, Stiegler A, King P, Boggon T. The C2 domain augments Ras GTPase-activating protein catalytic activity. Proceedings Of The National Academy Of Sciences Of The United States Of America 2025, 122: e2418433122. PMID: 39899710, PMCID: PMC11831179, DOI: 10.1073/pnas.2418433122.Peer-Reviewed Original ResearchConceptsGTPase-activating proteinC2 domainActivity of GTPase-activating proteinGTPase-activating protein domainProtein catalytic activityDomain in vitroAllosteric lobeRas GTPasesSequence conservationGTPase activityAlphaFold modelsRasGAPSignaling defectsRasMutationsCatalytic activityConstitutive disruptionCatalytic advantageGTPaseAlphaFoldDomainGenesSynGAPProteinSequence
2024
120 Treatable Acute Neuroinflammatory Disease Associated with Complement Factor I Loss-of-function in the Plain Community
Reid W, Carson V, Krieger P, Stiegler A, Boggon T, Sullivan K, van den Heuvel L, Romberg N. 120 Treatable Acute Neuroinflammatory Disease Associated with Complement Factor I Loss-of-function in the Plain Community. Clinical Immunology 2024, 262: 110062. DOI: 10.1016/j.clim.2024.110062.Peer-Reviewed Original Research
2023
Structure Determination of SH2–Phosphopeptide Complexes by X-Ray Crystallography: The Example of p120RasGAP
Stiegler A, Boggon T. Structure Determination of SH2–Phosphopeptide Complexes by X-Ray Crystallography: The Example of p120RasGAP. Methods In Molecular Biology 2023, 2705: 77-89. PMID: 37668970, PMCID: PMC11059313, DOI: 10.1007/978-1-0716-3393-9_5.Peer-Reviewed Original ResearchConceptsSrc homology 2SH2 domain bindsSH2 domainDomain bindsNew molecular-level insightsSH2 domain proteinsMolecular-level insightsX-ray crystallographyX-ray diffraction studiesDomain proteinsPartner proteinsHomology 2Three-dimensional structureMolecular detailsStructure determinationSuitable crystalsCanonical interactionsVapour-diffusion methodCareful structural analysisDrop vapor diffusion methodCrystallographic studiesCrystallography studiesSH2-phosphopeptide complexesDiffraction studiesP120RasGAPDiverse p120RasGAP interactions with doubly phosphorylated partners EphB4, p190RhoGAP, and Dok1
Vish K, Stiegler A, Boggon T. Diverse p120RasGAP interactions with doubly phosphorylated partners EphB4, p190RhoGAP, and Dok1. Journal Of Biological Chemistry 2023, 299: 105098. PMID: 37507023, PMCID: PMC10470053, DOI: 10.1016/j.jbc.2023.105098.Peer-Reviewed Original ResearchConceptsSH2 domainSpatial-temporal regulationDual SH2 domainsProper vascular developmentKey binding partnerProtein familySH2 interactionsBinding partnerHuman proteinsDistinct binding interactionsWeakened affinityVascular developmentRasGAPConformational differencesP190RhoGAPSmall-angle X-ray scatteringBindingBinding interactionsAffinity measurementsEphB4DomainGTPaseDok1X-ray scatteringProteinDe novo variants implicate chromatin modification, transcriptional regulation, and retinoic acid signaling in syndromic craniosynostosis
Timberlake A, McGee S, Allington G, Kiziltug E, Wolfe E, Stiegler A, Boggon T, Sanyoura M, Morrow M, Wenger T, Fernandes E, Caluseriu O, Persing J, Jin S, Lifton R, Kahle K, Kruszka P. De novo variants implicate chromatin modification, transcriptional regulation, and retinoic acid signaling in syndromic craniosynostosis. American Journal Of Human Genetics 2023, 110: 846-862. PMID: 37086723, PMCID: PMC10183468, DOI: 10.1016/j.ajhg.2023.03.017.Peer-Reviewed Original ResearchConceptsDamaging de novo variantsChromatin modificationsDe novo variantsCranial neural crest cellsGenome-wide significanceNeural crest cellsNovo variantsRetinoic acid receptor alphaExome sequence dataAcid receptor alphaTranscriptional regulationProband-parent triosGene transcriptionSequence dataCrest cellsOsteoblast differentiationCS phenotypeMendelian formsRecurrent gainsGenesRisk genesGenetic etiologyRetinoic acidReceptor alphaNeurodevelopmental disorders