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Tyrosine phosphoryl...
Tyrosine phosphorylation profiling via in situ proximity ligation assay
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- Elfineh, Lioudmila (författare)
- Uppsala universitet,Cancer och vaskulärbiologi,Science for Life Laboratory, SciLifeLab
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- Classon, Christina (författare)
- Uppsala universitet,Molekylära verktyg,Science for Life Laboratory, SciLifeLab
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- Asplund, Anna (författare)
- Uppsala universitet,Molekylär och morfologisk patologi,Science for Life Laboratory, SciLifeLab
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- Pettersson, Ulf (författare)
- Uppsala universitet,Genomik,Science for Life Laboratory, SciLifeLab
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- Kamali-Moghaddam, Masood (författare)
- Uppsala universitet,Molekylära verktyg,Science for Life Laboratory, SciLifeLab
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- Lind, Sara Bergström (författare)
- Uppsala universitet,Analytisk kemi,Institutionen för immunologi, genetik och patologi,Science for Life Laboratory, SciLifeLab
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(creator_code:org_t)
- 2014-06-13
- 2014
- Engelska.
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Ingår i: BMC Cancer. - : Springer Science and Business Media LLC. - 1471-2407. ; 14, s. 435-
- Relaterad länk:
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https://uu.diva-port... (primary) (Raw object)
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https://bmccancer.bi...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- Background: Tyrosine phosphorylation (pTyr) is an important cancer relevant posttranslational modification since it regulates protein activity and cellular localization. By controlling cell growth and differentiation it plays an important role in tumor development. This paper describes a novel approach for detection and visualization of a panel of pTyr proteins in tumors using in situ proximity ligation assay. Methods: K562 leukemia cells were treated with tyrosine kinase and/or phosphatase inhibitors to induce differences in pTyr levels and mimic cells with different malignant properties. Cells were then probed with one antibody against the pTyr modification and another probe against the detected protein, resulting in a detectable fluorescent signal once the probes were in proximity. Results: Total and protein specific pTyr levels on ABL, SHC, ERK2 and PI3K proteins were detected and samples of control and treated cells were distinguished at the pTyr level using this novel approach. Promising results were also detected for formalin fixed and paraffin embedded cells in the micro array format. Conclusions: This application of in situ proximity ligation assay is valuable in order to study the pTyr modification of a panel of proteins in large data sets to validate mass spectrometric data and to be combined with tissue microarrays. The approach offers new opportunities to reveal the pTyr signatures in cells of different malignant properties that can be used as biomarker of disease in the future.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Klinisk medicin -- Cancer och onkologi (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Clinical Medicine -- Cancer and Oncology (hsv//eng)
Nyckelord
- Cancer biomarkers
- Protein signaling
- Protein tyrosine phosphorylation
- in situ proximity ligation assay (in situ PLA)
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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