A mass spectrometry screening method for antiaggregatory activity of proteins covalently modified by combinatorial library members: application to sickle hemoglobin.
A homogeneous assay, based on electrospray mass spectrometry, is described for identifying compounds in a combinatorial library that covalently modify a protein and thereby enhance its solubility. The technique is based on measuring the distribution of modified proteins in the supernatant versus aggregate. Compounds having the greatest anti-aggregatory activity are those with the highest supernatant/aggregate ratio. Mass is used as a marker to identify which covalent modifier in the library is involved. An exploratory study is presented which demonstrates that the antisickling activity of a family of isothiocyanates, as measured by the standard C(sat) assay, correlates well (r(2) = 0.98) with the mass spectrometry analysis of the supernatant/aggregate distribution. The technique has potential for screening libraries capable of covalently modifying other proteins of clinical interest, e.g., Alzheimer's, Huntington's, and various prion related diseases.
Park
S
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Department of Medicinal Chemistry and Pharmacognosy and Center for Pharmaceutical Biotechnology, College of Pharmacy, University of Illinois at Chicago, 833 South Wood Street, Chicago, Illinois 60612, USA.
Wanna
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L
Johnson
M E
ME
Venton
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DL
D000986:Antisickling Agents / Q000138:chemical synthesis* / Q000737:chemistry / Q000494:pharmacology; D015195:Drug Design; D006451:Hemoglobin, Sickle / Q000737:chemistry*; D006801:Humans; D017879:Isothiocyanates / Q000138:chemical synthesis* / Q000737:chemistry* / Q000494:pharmacology; D013058:Mass Spectrometry / Q000379:methods; D013329:Structure-Activity Relationship
D000986:Antisickling Agents; D006451:Hemoglobin, Sickle; D017879:Isothiocyanates