Academic Journal

Decoding Arabidopsis thaliana CPK/SnRK Superfamily Kinase Client Signaling Networks Using Peptide Library and Mass Spectrometry

التفاصيل البيبلوغرافية
العنوان: Decoding Arabidopsis thaliana CPK/SnRK Superfamily Kinase Client Signaling Networks Using Peptide Library and Mass Spectrometry
المؤلفون: Nagib Ahsan, Amr R. A. Kataya, R. Shyama Prasad Rao, Kirby N. Swatek, Rashaun S. Wilson, Louis J. Meyer, Alejandro Tovar-Mendez, Severin Stevenson, Justyna Maszkowska, Grazyna Dobrowolska, Qiuming Yao, Dong Xu, Jay J. Thelen
المصدر: Plants, Vol 13, Iss 11, p 1481 (2024)
بيانات النشر: MDPI AG, 2024.
سنة النشر: 2024
المجموعة: LCC:Botany
مصطلحات موضوعية: kinase client assay, kinase substrates, mass spectrometry, phosphorylation, signaling network, Botany, QK1-989
الوصف: Members of the calcium-dependent protein kinase (CDPK/CPK) and SNF-related protein kinase (SnRK) superfamilies are commonly found in plants and some protists. Our knowledge of client specificity of the members of this superfamily is fragmentary. As this family is represented by over 30 members in Arabidopsis thaliana, the identification of kinase-specific and overlapping client relationships is crucial to our understanding the nuances of this large family of kinases as directed towards signal transduction pathways. Herein, we used the kinase client (KiC) assay—a relative, quantitative, high-throughput mass spectrometry-based in vitro phosphorylation assay—to identify and characterize potential CPK/SnRK targets of Arabidopsis. Eight CPKs (1, 3, 6, 8, 17, 24, 28, and 32), four SnRKs (subclass 1 and 2), and PPCK1 and PPCK2 were screened against a synthetic peptide library that contains 2095 peptides and 2661 known phosphorylation sites. A total of 625 in vitro phosphorylation sites corresponding to 203 non-redundant proteins were identified. The most promiscuous kinase, CPK17, had 105 candidate target proteins, many of which had already been discovered. Sequence analysis of the identified phosphopeptides revealed four motifs: LxRxxS, RxxSxxR, RxxS, and LxxxxS, that were significantly enriched among CPK/SnRK clients. The results provide insight into both CPK- and SnRK-specific and overlapping signaling network architectures and recapitulate many known in vivo relationships validating this large-scale approach towards discovering kinase targets.
نوع الوثيقة: article
وصف الملف: electronic resource
اللغة: English
تدمد: 2223-7747
Relation: https://www.mdpi.com/2223-7747/13/11/1481; https://doaj.org/toc/2223-7747
DOI: 10.3390/plants13111481
URL الوصول: https://doaj.org/article/dbae7624faab4a3ba3388eb033c69536
رقم الانضمام: edsdoj.bae7624faab4a3ba3388eb033c69536
قاعدة البيانات: Directory of Open Access Journals
الوصف
تدمد:22237747
DOI:10.3390/plants13111481