Recombinant Human DYRK1B, GST-tagged

• Cat.No.: DYRK1B-12237H
• Product Overview: Recombinant Human DYRK1B protein, fused to GST-tag, was expressed in E.coli and purified by GSH-sepharose.

Specification

• Species: Human
• Source: E.coli
• Tag: GST
• Protein Length: 111-431a.a.
• Description: DYRK1B is a member of the DYRK family of protein kinases. DYRK1B contains a bipartite nuclear localization signal and is found mainly in muscle and testis. The protein is proposed to be involved in the regulation of nuclear functions. Three isoforms of DYRK1B have been identified differing in the presence of two alternatively spliced exons within the catalytic domain.
• Storage: The protein is stored in PBS buffer at -20℃. Avoid repeated freezing and thawing cycles.
• Storage Buffer: 1M PBS (58mM Na2HPO4,17mM NaH2PO4, 68mM NaCl, pH8. ) added with 100mM GSH and 1% Triton X-100,15%glycerol.

Gene Information

• Gene Name: DYRK1B dual-specificity tyrosine-(Y)-phosphorylation regulated kinase 1B [ Homo sapiens ]
• Official Symbol: DYRK1B
• Synonyms: DYRK1B; dual-specificity tyrosine-(Y)-phosphorylation regulated kinase 1B; dual specificity tyrosine-phosphorylation-regulated kinase 1B; minibrain related kinase; MIRK; mirk protein kinase; minibrain-related kinase
• Gene ID: 9149
• mRNA Refseq: NM_004714
• Protein Refseq: NP_004705
• MIM: 604556
• UniProt ID: Q9Y463
• Chromosome Location: 19q12-q13.1
• Function: ATP binding; nucleotide binding; protein binding; protein kinase activity; protein serine/threonine kinase activity; protein serine/threonine/tyrosine kinase activity; protein tyrosine kinase activity; transcription coactivator activity

Citation

Dyrk1b promotes hepatic lipogenesis by bypassing canonical insulin signaling and directly activating mTORC2 in mice

Journal: The Journal of Clinical Investigation    PubMed ID: 34855620    Data: 2022/2/1

Authors: Neha Bhat, Anand Narayanan, Arya Mani

Article Snippet:All incubations at 4°C were done on an end-to-end rotator.All incubations at 4°C were done on an end-to-end rotator.. For the kinase reactions involving purified proteins, the immunoprecipitated mTORC2 or IgG was incubated with purified recombinant human DYRK1B (rhDYRK1B; Creative BioMart, DYRK1B-2983H, 81% pure) in the KRB buffer plus 500 μM ATP.. To pharmacologically inhibit DYRK1B, the rhDYRK1B was preincubated in either DMSO or AZ191 at a concentration of 10 μM for 10 minutes at room temperature.To pharmacologically inhibit DYRK1B, the rhDYRK1B was preincubated in either DMSO or AZ191 at a concentration of 10 μM for 10 minutes at room temperature.

( A ) Dyrk1b transcripts normalized to Hprt in the indicated conditions. Henceforth, each dot in the dot plots represents biological replicates; n > 8 mice each, unpaired t test, 2-tailed. ( B and C ) Western blot (WB) analysis and quantification of specified liver proteins in mice fed CD or HCD. Henceforth, each lane of WBs represents biological replicates; n = 4 mice each, unpaired t test, 2-tailed. ( D ) Correlation analysis between NASH score and Dyrk1b expression in the liver of HCD-fed mice. ( E ) WB and quantification of Dyrk1b levels in the nuclear (top) and cytoplasmic extracts (bottom) in the liver of mice fed with CD or HCD; n > 6 each, unpaired t test, 2-tailed. ( F and G ) Dyrk1b protein expression visualized by immunofluorescence in the liver of CD- or HCD-fed mice. IgG was used as control (top row); n = 5 mice each group. Arrows indicate cytoplasmic expression. Unpaired t test, 2-tailed. ( H and I ) Representative images and quantification of DYRK1B expression in the liver biopsies of patients with NASH versus controls; n = 20 controls, n = 27 NASH samples, unpaired t test, 2-tailed, Welch-corrected. ( J ) Correlation analysis between NASH score and percentage Dyrk1b expression in the human NASH samples ( n = 22) from I . Scale bars: 20 μm. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001.

( A ) Schematic representation of generation of AAV8 for increasing Dyrk1b levels in the mouse liver. ( B – E ) Oil Red O (ORO) staining, counterstained with hematoxylin ( B – D ) and total hepatic TAG ( E ) in the designated mice on CD after 6-hour fast; n > 6 each, 1-way ANOVA, Tukey’s post hoc test. Arrows in B – D indicate neutral lipid staining. ( F ) Fasting plasma TAG of the indicated mice on CD after 6-hour fast; n > 6 mice each, 1-way ANOVA, Tukey’s post hoc test. ( G ) Fasting TC on CD after 6-hour fast; n > 5 mice each, 1-way ANOVA, Tukey’s post hoc test.* P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001. Scale bar: 150 μm.

( A – H ) Representative confocal images from littermate controls and Dyrk1b –/– hepatocytes at indicated time points after addition of BODIPY-FA; n = 5 mice each genotype, n = 4 technical replicates. ( I ) Quantification of intracellular fluorescence in controls and Dyrk1b –/– hepatocytes measured by the microplate reader after cell lysis at the indicated time points. The background fluorescence (prior to addition of BODIPY-FA) was subtracted and normalized to the total protein content. Unpaired t test, 2-sided, n = 5 mice each genotype, n = 4 technical replicates. ( J – L ) Representative confocal images showing BODIPY-FA uptake 1 minute after addition in Dyrk1b –/– hepatocytes transduced with AAV8 containing empty vector (AAV control ) or Dyrk1b WT or Dyrk1b kin.def virus, at an MOI of 60. The FA uptake experiment was performed 72 hours after virus transduction, to allow for sufficient transcription. n = 2 mice each condition, n = 4 technical replicates. ( M ) Quantification of intracellular fluorescence, in the indicated genotypes; 1-way ANOVA, Tukey’s post hoc test. White arrows indicate the intracellular fluorescence detected by BODIPY-FA. Scale bars: 150 μm. ** P ≤ 0.01, *** P ≤ 0.001.