Identification of cell proliferation-associated epitope on CD98 oncoprotein using phage display random peptide library.
CD98 is known as a cell surface antigen expressed in proliferating normal tissues and in almost all tumor cells. Although the function of CD98 is not yet fully elucidated, it is suggested that CD98 is concerned functionally in lymphocyte activation, cell proliferation, and malignant transformation. Monoclonal antibody against human CD98 heavy chain (h.c.), termed HBJ127, shows inhibition of lymphocyte activation and tumor cell growth in vitro. These observations suggest that the epitope recognized by HBJ127 may be crucial for CD98 function. In the present study, the authors investigated the epitope recognized by HBJ127 using a phage display random heptapeptide library. Approximately 2.4 x 10(4)-fold amplification of eluted phage titer was obtained after three rounds of panning of the phage library against HBJ127. Seven different heptapeptide sequences were isolated from 30 randomly selected clones of the post-panning phage population. A homology search using ClustalW identified the peptide sequence corresponding to (442)AFS(444) of human CD98 h.c. It was also found that (443)F is a human-specific amino acid by comparing sequences of human, rat, and mouse origin. Reduced reactivity of HBJ127 was detected against the phenylalanine-substituted peptide but not detected against the alanine or serine-substituted one. It has been identified that HBJ127 reacts only with human species and the HBJ127 epitope position is predicted in 418-529 of human CD98 h.c. From these results and observations, it was estimated that (442)AFS(444) of human CD98 h.c. may be the HBJ127 epitope. Moreover, (443)F may be critical for the binding of HBJ127 against human CD98 h.c.
Itoh
Kunihiko
K
Department of Clinical Pharmacology and Genetics, School of Pharmaceutical Sciences, University of Shizuoka, 52-1 Yada, Suruga-ku, Shizuoka 422-8526, Japan. itohk@u-shizuoka-ken.ac.jp
Inoue
Kazuyuki
K
Hayashi
Hideki
H
Suzuki
Toshio
T
Masuko
Takashi
T
D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000737:chemistry / Q000235:genetics*; D001435:Bacteriophages; D002455:Cell Division; D004277:DNA, Single-Stranded / Q000737:chemistry / Q000235:genetics; D000939:Epitopes / Q000235:genetics; D027261:Fusion Regulatory Protein-1 / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D009363:Neoplasm Proteins / Q000235:genetics; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000737:chemistry; D019151:Peptide Library
D015703:Antigens, CD; D004277:DNA, Single-Stranded; D000939:Epitopes; D027261:Fusion Regulatory Protein-1; D009363:Neoplasm Proteins; D010446:Peptide Fragments; D019151:Peptide Library
J Mol Biol. 1995 Apr 14;247(5):932-46
7536850
Nat Biotechnol. 2005 Sep;23(9):1105-16
16151404
Cancer Res. 2007 Apr 15;67(8):3518-23
17440057
J Natl Cancer Inst. 2005 Nov 16;97(22):1663-70
16288119
J Mol Biol. 1992 Oct 5;227(3):711-8
1404385
Gan. 1983 Dec;74(6):819-21
6607858
Biochem Biophys Res Commun. 1999 Sep 7;262(3):720-5
10471392
Biochemistry. 1995 Mar 28;34(12):3948-55
7535098
J Biol Chem. 1998 Sep 11;273(37):23629-32
9726963
Biochem J. 1995 Dec 15;312 ( Pt 3):863-70
8554532
Oncogene. 2000 Dec 14;19(54):6209-15
11175335
J Biochem. 1999 Jan;125(1):123-9
9880807
Cancer Res. 1986 Mar;46(3):1478-84
2417704
Gene. 1993 Jun 15;128(1):29-36
8508957
Eur J Immunol. 2006 May;36(5):1267-74
16568495
J Immunol. 1981 Apr;126(4):1409-14
7204970
Jpn J Cancer Res. 1989 Jan;80(1):10-4
2565325
J Immunol. 1986 Mar 15;136(6):2055-61
3950410
Adv Immunol. 1994;57:191-280
7872158
Science. 1985 Jun 14;228(4705):1315-7
4001944
Annu Rev Immunol. 1994;12:433-55
8011287
Methods Mol Biol. 2002;178:1-37
11968478
Int Immunol. 2007 Mar;19(3):331-6
17272286
Cell Immunol. 2006 Feb;239(2):136-43
16814270
Jpn J Cancer Res. 1985 May;76(5):386-94
3924712
Eur J Immunol. 1994 Nov;24(11):2868-73
7525309
Cancer Res. 2005 Apr 15;65(8):3410-8
15833876
J Immunol. 2004 Jul 1;173(1):394-401
15210798
J Biol Chem. 1994 Sep 30;269(39):23853-6
7929027