1998 Fiscal Year Final Research Report Summary
Analysis of Lysosomal Protein Targeting Signals.
| Project/Area Number |
09670140
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General medical chemistry
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| Research Institution | Okayama University of Science |
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Principal Investigator |
NISHIKAWA Atsushi Okayama University of Science, Department of Science, Assistant Professor, 理学部, 講師 (30218127)
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| Project Period (FY) |
1997 – 1998
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| Keywords | Lysosome / Transportation / Signal |
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| Research Abstract |
We have reported that bovine DNase I, a secretary glycoprotein, acquires mannose 6-phosphate residues on 12.6% of its Asn-linked oligosaceharides when expressed in COS-1 cells and that the extent of phosphorylation increase to 79.2% when lysine are replaced at positions at 27 and 74 of the mature protein (Nishikawa, et al. (1997) J.Biol. Chem. 272, 19408-19412). We now demonstrate that muiine DNase I, which contains Lys^<27> and Lys74, is only phosphorylated 20.9% when expressed in the same COS-1 cell system. The difference is mostly due to the presence of three residues in murine sequence (Val^<23>, Lys^<117>, and Pro^<190>) that inhibit phosphorylation. Replacement of these residues with ahnines resulted in a strong stimulation of phosphorylation. In addition, substitution oh two other residues in the mouse sequence with the equivalent residues present in bovine DNase I (Glu57Tyr and Glul24Lys), but not replacement of these residues with an alanine, also resulted in enhanced phosphorylation, suggesting that these bovine residues have a positive stimulatory effect. The quadruple mutant (Lys117Ala-Prol 9OAla-Glu54Tyr-Val23Ala) was 65% phosphorylated, almost equivalent to the level obtained with bovine DNase I containing Lys^<27> and Lys^<74>.
These results indicate that the conformation-dependent recognition domain on murine DNase I that serves as the binding site for UDP-GIcNAc : Lysosomal enzyme NU-acetylglucosamine- l -phosphotransferase is suppressed by several inhibitory amino acid. In addition, murme DNase I lacks two of the stimulatory amino acids present in bovine DNase I, inducing a critical tyrosine residues.
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Research Products
(8 results)
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[Publications] Tanemura, M., Miyazawa, S.Ihara, Y., Nishikawa, A., Suzuki, M., Yamamura, K., Mastuda, H., Shirakura, R., and Taniguchi N.: ""Suppression of the xenoantigen Gal alpha(1,3)Gal by N-acetylglucosaminyl-transferase III(GnT-III)in transgenic mice"" Transplant Proc.29. 895-896 (1997)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Ihara, Y., Yoshimura, M., Miyoshi, E., Nishikawa, A., Sultan, A.S., Toyosawa, S., Ohnishi, A., Suzuki, M., yamamura, K., Ijyuhin N., and Taniguchi N.: ""Ectopic expression of N-acetylglucosaminyltransferase III in transgenic hepatocytes disrupts apolipoprotein B secretion and induces aberrant cellular morphology with lipid storage"" Proc.Natl.Acad.Sci.95. 2526-2530 (1998)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Taniguchi, N., Yoshimura, M., Miyoshi, E., Ihara, Y., Nishikawa, A., Kang R.and Ikeda Y.: ""Gene expression and regulation of N-acetylglucosaminyltransferases III and V in cancer tissues"" Advan.Enzyme Regul.38. 223-232 (1998)
Description
「研究成果報告書概要(欧文)」より
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