Screening proteins interacting with hCLP46 using tandem affinity purification

doi:10.7523/j.issn.2095-6134.2011.2.011

Abstract

Abstract:

hCLP46 (human CAP10-like protein46), a novel gene, has been screened out from the cDNA library of MDS-AML Patient’s CD34⁺ stem cell. For to further studying the biological characteristics of this gene tandem affinity purification (TAP) has been used to isolate proteins which specifically interact with hCLP46, and seven specific protein bands were detected. By using liquid chromatography-mass spectrometry and protein database searching, a series of endoplasmic reticulum chaperones were identified. Two major ER chaperone systems, the BiP/Grp94 and the calnexin (CNX)/ calreticulin (CRT) systems, are important in the maturation of hCLP46.

Key words: crosslinker, tandem affinity purification(TAP), MDS, hCLP46

CLC Number:

XU Feng, MU Xin, WANG Wei, LIU Li-Xin. Screening proteins interacting with hCLP46 using tandem affinity purification[J]. , 2011, 28(2): 210-216.

References

[1] Teng Y, Liu Q, Ma J, et al. Cloning, expression and characterization of a novel human CAP10-like gene hCLP 46 from CD34+ stem/progenitor cells
[J]. Gene, 2006, 371: 7-15.

[2] Ponting C P, Mott R, Bork P, et al. Novel protein domains and repeats in Drosophila melanogaster: Insights into structure, function, and evolution
[J]. Genome Research, 2001, 11: 1996-2008.

[3] Acar M, Jafar-Nejad H, Takeuchi H, et al. Rumi is a CAP10 domain glycosyltransferase that modifies notch and is required for notch signaling
[J]. Cell, 2008, 132: 247-258.

[4] Zanotti S, Canalis E. Notch and the Skeleton
[J]. Molecular and Cellular Biology, 2010, 30: 886-896.

[5] Rigaut G, Shevchenko A, Rutz B, et al. A generic protein purification method for protein complex characterization and proteome exploration
[J]. Nature Biotechnology, 1999, 17: 1030-1032.

[6] Goto H, Kawaoka Y. A novel mechanism for the acquisition of virulence by a human influenza A virus
[J]. Proceedings of the National Academy of Sciences of the United States of America, 1998, 95: 10224-10228.

[7] Haas I G, Wabl M. Immunoglobulin heavy chain binding protein
[J]. Nature, 1983, 306: 387-389.

[8] Lievremont J P, Rizzuto R, Hendershot L, et al. BiP, a major chaperone protein of the endoplasmic reticulum lumen, plays a direct and important role in the storage of the rapidly exchanging pool of Ca²⁺
[J]. J Biol Chem, 1997, 272: 30873-30879.

[9] Kabani M, Kelley S S, Morrow M W, et al. Dependence of endoplasmic reticulum-associated degradation on the peptide binding domain and concentration of BiP
[J]. Mol Biol Cell, 2003, 14: 3437-3448.

[10] Molinari M, Helenius A. Chaperone selection during glycoprotein translocation into the endoplasmic reticulum
[J]. Science, 2000, 288: 331-333.

[11] Ellgaard L, Helenius A. ER quality control: towards an understanding at the molecular level
[J]. Current Opinion in Cell Biology, 2001, 13: 431-437.

[12] Hebert D N, Molinari M. In and out of the ER: Protein folding, quality control, degradation, and related human diseases
[J]. Physiological Reviews, 2007, 87: 1377-1408.

[13] Helenius A, Aebi M. Roles of N-linked glycans in the endoplasmic reticulum
[J]. Annu Rev Biochem, 2004, 73: 1019-1049.

[14] Trombetta E S, Simons J F, Helenius A. Endoplasmic reticulum glucosidase II is composed of a catalytic subunit, conserved from yeast to mammals, and a tightly bound noncatalytic HDEL-containing subunit
[J]. Journal of Biological Chemistry, 1996, 271: 27509-27516.

[15] Chevet E, Jakob C A, Thomas D Y, et al. Calnexin family members as modulators of genetic diseases
[J]. Seminars in Cell and Developmental Biology, 1999, 10: 473-480.

[16] Cai H, Wang C C, Tsou C L. Chaperone-like activity of protein disulfide-isomerase in the refolding of a protein with no disulfide bonds
[J]. Journal of Biological Chemistry, 1994, 269: 24550-24552.

[17] Quan H, Fan G B, Wang C C. Independence of the chaperone activity of protein disulfide-isomerase from its thioredoxin-like active-site
[J]. Journal of Biological Chemistry, 1995, 270: 17078-17080.

[18] Maattanen P, Kozlov G, Gehring K, et al. ERp57 and PDI: multifunctional protein disulfide isomerases with similar domain architectures but differing substrate-partner associations
[J]. Biochemistry and Cell Biology-Biochimie Et Biologie Cellulaire, 2006, 84: 881-889.

[19] Oliver J D, Roderick H L, Llewellyn D H, et al. ERp57 functions as a subunit of specific complexes formed with the ER lectins calreticulin and calnexin
[J]. Molecular Biology of the Cell, 1999, 10: 2573-2582.

[20] Chevet E, Wong H N, Gerber D, et al. Phosphorylation by CK2 and MAPK enhances calnexin association with ribosomes
[J]. Embo Journal, 1999, 18: 3655-3666.