biochemical evidence for a novel low molecular weight 2-5a.pdfVIP

JOURNAL OF INTERFERON AND CYTOKINE RESEARCH 17:377-385 (1997) Mary Ann Liebert, Inc. Biochemical Evidence for a Novel Low Molecular Weight 2-5A-Dependent RNase L in Chronic Fatigue Syndrome 1,2 3 3 ROBERT I SUHADOLNIK, DANIEL L PETERSON, KAREN OBRIEN, PAUL 4 1 1 1 R. CHENEY, C.V.T. HERST, NANCY L. REICHENBACH, NING KON, SUSAN 1 2 2 E. HORVATH, KATHRYN T. IACONO, MARTIN E. ADELSON, KENNY DE 5 5 6 MEIRLEIR, PASCALE DE BECKER, RAMAMURTHY CHARUBALA, and WOLFGANG PFLEIDERER6 ABSTRACT Previous studies from this laboratory have demonstrated a statistically significant dysregulation in several key components of the 2,5-oligoadenylate (2-5A) synthetaseIRNase Land PKR antiviral pathways in chronic fatigue syndrome (CFS) (Suhadolnik et al. Clin Infect Dis 18, 596104, 1994; Suhadolnik et al. In Vivo 8, 599~04, 1994). Two methodologies have been developed to further examine the upregulated RNase L activity in CFS. First, photoaffinity labeling of extracts of peripheral blood mononuclear cells (PBMC) with the azido 2-5A photoaffinity probe, [32p ]ApAp(8-azidoA), followed by immunoprecipitation with a polyclonal antibody against recombinant, human 80-kDa RNase L and analysis under denaturing conditions. A subset of individuals with CFS was identified with only one 2-5A binding protein at 37 kDa, whereas in extracts of PBMC from a second subset of CFS PBMC and from healthy controls, photolabeled/limmunoreactive 2-5A binding proteins were detected at 80, 42, and 37 kl)a. Second, analytic gel permeati