Over the 30 years'duration of this grant, this project has established a broad foundation for understanding the role of carbohydrate structures on glycoproteins. In this time our studies have defined 1) unique oligosaccharide structures as signatures found on specific plasma glycoproteins, 2) the mechanisms of recognition of these structures by carbohydrate-specific endocytic receptors, and 3) the functionally important in vivo biological consequences of this recognition. We have proceeded from determining at the molecular and cellular level the biologic basis of oligosaccharide recognition by these receptors, to delineating at the whole organism level the functional implications of such recognition. Our progress on how carbohydrate-specific endocytic receptor recognition systems clear particular glycoproteins from the circulation now culminates in a unique opportunity to take these studies to a new level of functional perspective. We have demonstrated by biochemical and genetic means that the Mannose/GalNAc-4-SO4-receptor (M/G4S-R) and the asialoglycoprotein-receptor (ASGP-R) mediate the respective clearance of glycoproteins that bear the N-linked oligosaccharide signatures of terminal GalNAc 4 SO4 and of terminal Sia12,6GalNAc. We have shown that the ASGP-R can bind glycoproteins bearing Sia12,6Gal. We hypothesize that glycoproteins bearing terminal Sia12,6Gal are in fact the major endogenous ligands for the ASGP-R and that the ASGP-R is a key regulator of the concentration of numerous plasma glycoproteins that have Sia12,6Gal termini. Preliminary studies support this proposed function. Our findings give a significant new functional identity to this receptor. We will now use genetic models and recently established proteomic methods to define the relative roles of the ASGP-R and M/G4S-R. We will determine how these receptors act, individually and in combination, to regulate levels of plasma glycoproteins based on their carbohydrate signatures, under physiologic as well as pathologic conditions ranging from regulation of estrogen production by pituitary glycoprotein hormones to the acute phase response to sepsis. These proposed studies utilize single and double receptor knockout mice 1) to address how production is balanced with clearance to regulate plasma levels of glycoproteins through recognition of their carbohydrate signatures, 2) to determine if perturbing this balance has an impact on biologic processes, and 3) to establish the molecular basis for generating isoforms of the M/G4S-R with differing carbohydrate signature specificities and determine the impact of such receptor isoforms on reproductive biology. The fundamental mechanistic information that we uncover will ultimately yield insights that can aid in evaluating changes in plasma proteins for both diagnostic and therapeutic purposes. Public Health Relevance: The levels of glycoproteins in the blood reflect a balance between their synthesis and removal. We have identified two major carbohydrate specific receptor systems in liver that help control the concentrations of many glycoproteins in the blood by determining their rates of removal. We will define how these receptors control the concentrations of hormones and inflammatory proteins during pregnancy and the response to injury.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA021923-34
Application #
8259178
Study Section
Special Emphasis Panel (ZRG1-ICI-D (01))
Program Officer
Snyderwine, Elizabeth G
Project Start
1977-08-01
Project End
2014-05-31
Budget Start
2012-06-01
Budget End
2014-05-31
Support Year
34
Fiscal Year
2012
Total Cost
$636,622
Indirect Cost
$217,792
Name
Washington University
Department
Pathology
Type
Schools of Medicine
DUNS #
068552207
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Mi, Yiling; Lin, Angela; Fiete, Dorothy et al. (2014) Modulation of mannose and asialoglycoprotein receptor expression determines glycoprotein hormone half-life at critical points in the reproductive cycle. J Biol Chem 289:12157-67
Fiete, D J; Beranek, M C; Baenziger, J U (1998) A cysteine-rich domain of the "mannose" receptor mediates GalNAc-4-SO4 binding. Proc Natl Acad Sci U S A 95:2089-93
Fiete, D; Beranek, M C; Baenziger, J U (1997) The macrophage/endothelial cell mannose receptor cDNA encodes a protein that binds oligosaccharides terminating with SO4-4-GalNAcbeta1,4GlcNAcbeta or Man at independent sites. Proc Natl Acad Sci U S A 94:11256-61
Fiete, D; Baenziger, J U (1997) Isolation of the SO4-4-GalNAcbeta1,4GlcNAcbeta1,2Manalpha-specific receptor from rat liver. J Biol Chem 272:14629-37
Manzella, S M; Hooper, L V; Baenziger, J U (1996) Oligosaccharides containing beta 1,4-linked N-acetylgalactosamine, a paradigm for protein-specific glycosylation. J Biol Chem 271:12117-20
Hooper, L V; Manzella, S M; Baenziger, J U (1996) From legumes to leukocytes: biological roles for sulfated carbohydrates. FASEB J 10:1137-46
Haslam, D B; Baenziger, J U (1996) Expression cloning of Forssman glycolipid synthetase: a novel member of the histo-blood group ABO gene family. Proc Natl Acad Sci U S A 93:10697-702
Musil, L S; Baenziger, J U (1987) Cleavage of membrane secretory component to soluble secretory component occurs on the cell surface of rat hepatocyte monolayers. J Cell Biol 104:1725-33
Colley, K J; Baenziger, J U (1987) Biosynthesis and secretion of the rat core-specific lectin. Relationship of post-translational modification and assembly to attainment of carbohydrate binding activity. J Biol Chem 262:3415-21
Green, E D; Boime, I; Baenziger, J U (1986) Differential processing of Asn-linked oligosaccharides on pituitary glycoprotein hormones: implications for biologic function. Mol Cell Biochem 72:81-100

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