Residualizing labels are biologically inert, non-degradable radioactive or fluorescent tags for proteins which are retained in lysosomes following cellular uptake and catabolism of the carrier protein. Our laboratory has been involved in the development of these labels for a number of years, beginning with (3H)raffinose in 1978, and followed by dilactitol-125I-tyramine in 1985. These residualizing labels have proven useful in identifying the tissue and cell sites of catabolism of long-lived plasma proteins such as albumin, lipoproteins and immunoglobulins in vivo, as well as in studies on the uptake and catabolism of proteins by cells in culture. During the last two years we have prepared and characterized an improved radioactive label, inulin-125I-tyramine, and a new fluorescent label, N,N--dilactitol-N'-fluoresceinyl- ethylenedlamine, and have continued studies on the catabolism of albumin and IgM in vivo and on the catabolism of albumin by fibroblasts in culture. We have also made progress toward development of an 18F-label for use in noninvasive imaging by positron emission tomography. Our research goals for the next five year period are a continuation of our current effort to develop more useful labels, to use them in studies on the sites and mechanisms of plasma protein catabolism and to collaborate in the application of these labels for noninvasive and diagnostic imaging.
Specific Aims are: 1. Continued work on fluorescent labels, to include development of a red fluorescent label and application of these labels in experiments in vivo and in vitro using fluorescence microscopy and fluorescence activated cell sorting; 2. Continued work on the synthesis and application of residualizing labels for non-invasive imaging, to include development of 18-F-labels suitable for positron emission tomography and 19F-Labels for magnetic resonance imaging, and application of these labels in studies on deposition of circulating lipoproteins, diagnostic imaging with anti-tumor antibodies and targeting of protein pharmaceuticals; and 3. Continued studies on the catabolism of serum proteins in vivo, to include (a) studies on the mechanisms involved in the regulation of albumin catabolism, with emphasis on the effects of fatty acid saturation on albumin catabolism, (b) studies on the sites of catabolism of IgG and IgM and the role of receptors in catabolism of these proteins, (c) studies on the catabolism of lactate dehydrogenase isozymes to determine the biochemical basis for differential kinetics of catabolism of the M4(short-lived) isozymes, and (d) studies on the catabolism of oxidized serum proteins.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK025373-12
Application #
3227354
Study Section
Special Emphasis Panel (SSS (B))
Project Start
1979-04-01
Project End
1993-07-31
Budget Start
1990-08-01
Budget End
1991-07-31
Support Year
12
Fiscal Year
1990
Total Cost
Indirect Cost
Name
University of South Carolina at Columbia
Department
Type
Schools of Arts and Sciences
DUNS #
111310249
City
Columbia
State
SC
Country
United States
Zip Code
29208
Stein, R; Goldenberg, D M; Thorpe, S R et al. (1995) Effects of radiolabeling monoclonal antibodies with a residualizing iodine radiolabel on the accretion of radioisotope in tumors. Cancer Res 55:3132-9
Chroneos, Z C; Baynes, J W; Thorpe, S R (1995) Identification of liver endothelial cells as the primary site of IgM catabolism in the rat. Arch Biochem Biophys 319:63-73
Thorpe, S R; Baynes, J W (1994) Residualizing glycoconjugates: biologically inert tracers for studies on protein endocytosis and catabolism. Methods Enzymol 242:3-17
Potter, D; Chroneos, Z C; Baynes, J W et al. (1993) In vivo fate of hemopexin and heme-hemopexin complexes in the rat. Arch Biochem Biophys 300:98-104
Thorpe, S R; Baynes, J W; Chroneos, Z C (1993) The design and application of residualizing labels for studies of protein catabolism. FASEB J 7:399-405
Wells-Knecht, M C; Huggins, T G; Dyer, D G et al. (1993) Oxidized amino acids in lens protein with age. Measurement of o-tyrosine and dityrosine in the aging human lens. J Biol Chem 268:12348-52
Bichler, J; Baynes, J W; Thorpe, S R (1993) Catabolism of hirudin and thrombin-hirudin complexes in the rat. Biochem J 296 ( Pt 3):771-6
Huggins, T G; Wells-Knecht, M C; Detorie, N A et al. (1993) Formation of o-tyrosine and dityrosine in proteins during radiolytic and metal-catalyzed oxidation. J Biol Chem 268:12341-7
Ord, J M; Hasapes, J; Daugherty, A et al. (1992) Imaging of thrombi with tissue-type plasminogen activator rendered enzymatically inactive and conjugated to a residualizing label. Circulation 85:288-97
Huggins, T G; Staton, M W; Dyer, D G et al. (1992) o-Tyrosine and dityrosine concentrations in oxidized proteins and lens proteins with age. Ann N Y Acad Sci 663:436-7

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