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Researchers gain fuller picture of cell protein reactions
Date:11/21/2013

processes called proplatelets and produce thousands of platelets, but in culture they produce fewer than 10 because the cells do not get as large and many die before they release platelets.

In an earlier study, Miller and his collaborators found that inhibiting a certain family of enzymes helped promote differentiation, resulting in larger megakaryocytes and more extensive proplatelet formation.

Miller's colleague, Milan Mrksich, the Henry Wade Rogers Professor of Biomedical Engineering, Chemistry, and Cell and Molecular Biology at McCormick, had been developing bioanalytical techniques for just this type of problem. The two partnered to profile a cell line model of the bone marrow cells that produce platelets.

"If we understand the enzyme activities that occur during megakaryocyte differentiation, it may be possible to prevent or promote differentiation for platelet production and other purposes," Miller said.

Using Mrksich's unique process of self-assembled monolayers desorption ionization (SAMDI) mass spectrometry, a super-fast, low-cost, and "label-free" method of measuring biochemical activities on a surface, the researchers were able to identify patterns of enzyme activities in cell lysates.

The researchers focused on histone deacetylase enzymes, a family of 17 enzymes that remove acetyl groups from certain proteins. They found that global deacetylase activity decreased significantly during differentiation, and that the decrease could be attributed to the sirtuin class comprising six deacetylases. The activities of the other 11 "classical" deacetylases did not substantially change.

Traditionally, discovering protein function has been a slow, tedious process of trial and error. Current methods use labels chemical additives that leave their mark in a reaction, such as radioactivity or fluorescence to determine whether a protein is active in a reaction. But labels can only test to see whether a specific r
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Contact: Erin White
ewhite@northwestern.edu
847-491-4888
Northwestern University
Source:Eurekalert

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