The envelope glycoprotein gp120 of the human immunodeficiency virus (HIV) is a high glycosylated molecule. It has been suggested that gp120 plays a vital role in viral attachment and the initiation of infection through interaction of CD4 glycoprotein of T-lymphocytes. This program centers at the chemical synthesis of oligosaccharide structures which are being reported as part of the carbohydrate moiety of gp120 and their further use as immunogens. Attention will be focussed on the synthesis of glycosides that possess an aglycon which can serve as a bridge for attachment to high molecular weight substances such as bovine serum albumin. Complex structures containing mannose is one of our major interests in this program. We plan to investigate the use of various derivatives of 4- pentenyl-alpha-D-mannopyranoside as glycosylating agents. The structures of our synthetic compounds will be established by n.m.r. studies and mass spectroscopy. Our synthetic antigens will be further utilized to raise their antibodies. The availability of antigen moiety linked to a solid support (Sepharose) will facilitate purification of the antibodies raised against the corresponding antigens. Specificity of the purified antibodies will be established with a battery of our well-defined carbohydrate structures. Upon purification the antibodies produced to the synthetic obligosaccharides will be tested for their ability to block gp120-CD4 binding and gp120-CD4 mediated cell fusion and for their ability to neutralize virus growth. Our synthetic saccharides linked to BSA will be used to develop saccharide ELISA methodology for the detection of anti- carbohydrate antibodies in the sera of AIDS patients. Thus, in the program we also plan to screen AIDS patients' sera for well-defined oligosaccharide specific antibodies. In another aspect of this program, we will emphasize our studies toward the chemical synthesis of various potential inhibitors of glycosidases, such as glucosidases and mannosidases, involved in the processing and biosynthesis of glycoprotein including gp120. The effect of glycosylation inhibitors on gp120 maturation will be assessed in a variety of transfected cell lines and in virus-infected cells. Our potential inhibitors may prove to be chemotherapeutic agents for AIDS.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI029326-02
Application #
3144086
Study Section
AIDS and Related Research Study Section 1 (ARRA)
Project Start
1991-09-01
Project End
1994-07-31
Budget Start
1992-09-01
Budget End
1993-07-31
Support Year
2
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Roswell Park Cancer Institute Corp
Department
Type
DUNS #
City
Buffalo
State
NY
Country
United States
Zip Code
14263
Chen, Y; Jain, R K; Chandrasekaran, E V et al. (1995) Use of sialylated or sulfated derivatives and acrylamide copolymers of Gal beta 1,3GalNAc alpha- and GalNAc alpha- to determine the specificities of blood group T- and Tn-specific lectins and the copolymers to measure anti-T and anti-Tn antibody levels in Glycoconj J 12:55-62
Chandrasekaran, E V; Jain, R K; Larsen, R D et al. (1995) Selectin ligands and tumor-associated carbohydrate structures: specificities of alpha 2,3-sialyltransferases in the assembly of 3'-sialyl-6-sialyl/sulfo Lewis a and x, 3'-sialyl-6'-sulfo Lewis x, and 3'-sialyl-6-sialyl/sulfo blood group T-hapten. Biochemistry 34:2925-36
Brockhausen, I; Reck, F; Kuhns, W et al. (1995) Substrate specificity and inhibition of UDP-GlcNAc:GlcNAc beta 1-2Man alpha 1-6R beta 1,6-N-acetylglucosaminyltransferase V using synthetic substrate analogues. Glycoconj J 12:371-9
Jain, R K; Liu, X G; Matta, K L (1995) Synthesis of isomeric sulfated disaccharides. Methyl O-(2-acetamido-2-deoxy-3-O-, 4-O-, and 6-O-sulfo-beta-D-glucopyranosyl sodium salt)-(1-->3)-beta-D-galactopyranoside. Carbohydr Res 268:279-85
Khan, S H; Matta, K L (1995) Synthesis of 2-acetamido-2-deoxy-beta-D-glucopyranosyl-(1-->2)-alpha-D- mannopyranosyl-(1-->6)-beta-D-mannopyranosyl-(1-->4)-2-acetamido-2-d eox y-D-glucopyranose. Acceptor-substrate recognition by N-acetylglucosaminyltransferase-V (GnT-V). Carbohydr Res 278:351-62
Kuhns, W; Jain, R K; Matta, K L et al. (1995) Characterization of a novel mucin sulphotransferase activity synthesizing sulphated O-glycan core 1,3-sulphate-Gal beta 1-3GalNAc alpha-R. Glycobiology 5:689-97
Jain, R K; Liu, X G; Oruganti, S R et al. (1995) Synthesis of oligosaccharide substrates for N-linked glycoprotein processing enzymes. Carbohydr Res 271:185-96
Chandrasekaran, E V; Jain, R K; Rhodes, J M et al. (1995) Expression of blood group Lewis b determinant from Lewis a: association of this novel alpha (1,2)-L-fucosylating activity with the Lewis type alpha (1,3/4)-L-fucosyltransferase. Biochemistry 34:4748-56
Powell, L D; Jain, R K; Matta, K L et al. (1995) Characterization of sialyloligosaccharide binding by recombinant soluble and native cell-associated CD22. Evidence for a minimal structural recognition motif and the potential importance of multisite binding. J Biol Chem 270:7523-32
Jain, R K; Piskorz, C F; Matta, K L (1995) Synthetic mucin fragments: synthesis of O-sulfo and O-methyl derivatives of allyl O-(beta-D-galactopyranosyl)-(1-->3)-2-acetamido-2-deoxy-alpha-D- galactopyranoside as potential compounds for sulfotransferases. Carbohydr Res 275:231-43

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