This renewal application has two related objectives. One is the detailed analysis by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) of lymphoid cells for the purpose of detecting polypeptides that are abnormally expressed in acute lymphoblastic leukemia (ALL). The analysis will benefit from the increased resolution of 2-D PAGE we have achieved, the development of computer programs for polypeptide spot matching and from the availability of a lymphoid protein database that we have established and that we propose to expand. The second objective consists of detailed investigations of a selected group of polypeptides of interest. We propose to focus initially on four polypeptides detected in ALL in the initial grant cycle, specifically: LT4 (T cell marker found in abundance in T ALL), L4 (common ALL marker), L3 (a polypeptide that is deficient in ALL subtypes associated with poor prognosis), and p18 (a polypeptide present in relatively high amount in ALL but not in other lymphoid proliferating cells examined). An immobilized pH gradient based approach, developed in this laboratory, will be relied upon to generate preparative amounts of polypeptides for sequencing and for monoclonal antibody production. Antibodies will be utilized to determine the occurrence of the corresponding polypeptides in single cells and for screening of samples using a simple one-dimensional electrophoresis technique. These studies are expected to result in a better understanding of the extent to which the pathogenetic process in leukemia is associated with abnormalities in a major constituent of cells through which function is mediated, namely the proteins.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
2R01CA032146-04A2
Application #
3170142
Study Section
Biochemistry Study Section (BIO)
Project Start
1983-02-01
Project End
1991-06-30
Budget Start
1988-07-01
Budget End
1989-06-30
Support Year
4
Fiscal Year
1988
Total Cost
Indirect Cost
Name
University of Michigan Ann Arbor
Department
Type
Schools of Medicine
DUNS #
791277940
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
Chang, C L; Hora, N; Huberman, N et al. (2001) Oncoprotein 18 levels and phosphorylation mediate megakaryocyte polyploidization in human erythroleukemia cells. Proteomics 1:1415-23
Melhem, R; Hailat, N; Kuick, R et al. (1997) Quantitative analysis of Op18 phosphorylation in childhood acute leukemia. Leukemia 11:1690-5
Chang, C L; Strahler, J R; Thoraval, D H et al. (1996) A nucleoside diphosphate kinase A (nm23-H1) serine 120-->glycine substitution in advanced stage neuroblastoma affects enzyme stability and alters protein-protein interaction. Oncogene 12:659-67
Ungar, D R; Hailat, N; Strahler, J R et al. (1994) Hsp27 expression in neuroblastoma: correlation with disease stage. J Natl Cancer Inst 86:780-4
Keim, D R; Hailat, N; Kuick, R et al. (1993) PCNA levels in neuroblastoma are increased in tumors with an amplified N-myc gene and in metastatic stage tumors. Clin Exp Metastasis 11:83-90
Strahler, J R; Zhu, X X; Hora, N et al. (1993) Maturation stage and proliferation-dependent expression of dUTPase in human T cells. Proc Natl Acad Sci U S A 90:4991-5
Wang, Y K; Liao, P C; Allison, J et al. (1993) Phorbol 12-myristate 13-acetate-induced phosphorylation of Op18 in Jurkat T cells. Identification of phosphorylation sites by matrix-assisted laser desorption ionization mass spectrometry. J Biol Chem 268:14269-77
Hanash, S M; Beretta, L; Barcroft, C L et al. (1993) Mapping of the gene for interferon-inducible dsRNA-dependent protein kinase to chromosome region 2p21-22: a site of rearrangements in myeloproliferative disorders. Genes Chromosomes Cancer 8:34-7
Hanash, S M; Strahler, J R; Chan, Y et al. (1993) Data base analysis of protein expression patterns during T-cell ontogeny and activation. Proc Natl Acad Sci U S A 90:3314-8
Gratiot-Deans, J; Keim, D; Strahler, J R et al. (1992) Differential expression of Op18 phosphoprotein during human thymocyte maturation. J Clin Invest 90:1576-81

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