Substrate specificity of soluble and membrane-associated ADP-ribosyltransferase ART2.1.
ADP-ribosyltransferases (ARTs) are a family of enzymes that catalyze the covalent transfer of an ADP-ribose moiety, derived from NAD, to an amino acid of an acceptor protein, thereby altering its function. To date, little information is available on the protein target specificity of different ART family members. ART2 is a T-cell-specific transferase, attached to the cell surface by a glycosylphosphatidylinositol (GPI) anchor, and also found in serum. Here we investigated the role of ART2 localization in serum or on the cell surface, or solubilized with detergents or enzymes, on its target protein specificity. We found that detergent solubilization of cell membranes, or release of ART2 by phosphoinositide-specific phospholipase C treatment, altered the ability of ART2 to ADP-ribosylate high or low molecular weight histone proteins. Similarly, soluble recombinant ART2 (lacking the GPI anchor) showed a different histone specificity than did cell-bound ART2. When soluble ART2 was incubated with serum proteins in the presence of [32P]-labeled NAD, several serum proteins were ADP-ribosylated in a thiol-specific manner. Mass spectrometry of labeled proteins identified albumin and transferrin as ADP-ribosylated proteins in serum. Collectively, these studies reveal that the membrane or solution environment of ART2 plays a pivotal role in determining its substrate specificity.
Zheng
Xuexiu
X
Department of Medicine, University of Massachusetts Medical School, Worcester, Massachusetts 01605, USA.
Morrison
Alan R
AR
Chung
An-Sik
AS
Moss
Joel
J
Bortell
Rita
R
D036002:ADP Ribose Transferases / Q000737:chemistry* / Q000235:genetics; D000244:Adenosine Diphosphate / Q000737:chemistry / Q000378:metabolism; D000818:Animals; D019556:COS Cells; D002462:Cell Membrane / Q000201:enzymology*; D002522:Chlorocebus aethiops; D051379:Mice; D009243:NAD / Q000737:chemistry / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012995:Solubility; D013379:Substrate Specificity / Q000502:physiology; D013601:T-Lymphocytes / Q000201:enzymology*
D011994:Recombinant Proteins; D009243:NAD; D000244:Adenosine Diphosphate; D036002:ADP Ribose Transferases; C449322:Art2a protein, mouse