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Contact spotting of protein microarrays coupled with spike‐in of normalizer protein permits time‐resolved analysis of ERBB receptor signaling
Author(s) -
Löbke Christian,
Laible Mark,
Rappl Claudia,
Ruschhaupt Markus,
Sahin Özgür,
Arlt Dorit,
Wiemann Stefan,
Poustka Annemarie,
Sültmann Holger,
Korf Ulrike
Publication year - 2008
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200700733
Subject(s) - erbb , neuregulin , epidermal growth factor receptor , erbb3 , protein microarray , signal transduction , computational biology , erbb4 , proteomics , biology , protein array analysis , receptor , microbiology and biotechnology , receptor tyrosine kinase , dna microarray , genetics , gene expression , gene
Abstract Protein microarrays allow highly accurate comparison and quantification of numerous biological samples in parallel while requiring only little material. This qualifies protein arrays for systems biology and clinical research where only limited sample material is available, but a precise readout is required. With the introduction of signal normalization steps to monitor the drop size of manually contact‐spotted RP protein arrays, the usefulness of normalizer proteins to ensure a high‐throughput but inexpensive protein analysis was demonstrated. This approach was applied for the analysis of signaling through ERBB receptor activated kinases in the breast cancer cell line MCF‐7. Activation of ERK1/2 and AKT by ERBB1 (EGFR), ERRB2 (HER2/neu), and ERBB3‐4 was monitored in a time‐resolved manner. Analysis of pathway activation by stimulation with epidermal growth factor and heregulin, or inhibition by blocking with gefitinib or herceptin allowed a characterization of the distinct signaling properties of the different ERBB receptor subtypes.