Artificial DnaJ Protein for Protein Production and Conformational Diseases

Overview

Journal Sci Rep

Specialty Science

Date 2017 Aug 19

PMID 28819167

Citations 2

Authors

Akinori Hishiya

Keizo Koya

Affiliations

Soon will be listed here.

Abstract

For secreted proteins, proper protein folding is essential not only for biological function but also for secretion itself. Proteins with folding problems are trapped in the endoplasmic reticulum (ER) and are eventually degraded in the cytoplasm. In this study, we exploited co-expression of an artificial fusion protein, based on the sequence of a DnaJ protein, which could interact as co-chaperones in the Hsp70-based protein-folding system, with target recombinant secreted proteins to enhance their production and secretion. The J-domain sequence or a fragment thereof was conjugated to a target protein-binding domain that was capable of binding to a portion of the target-protein sequence. Production of many of the target proteins was significantly upregulated when co-expressed with the J-domain fusion protein. Surprisingly, the enhancement of secretion was observed even when the J-domain had a mutation in the HPD motif, which is necessary for J-protein-Hsp70 interactions, suggesting the phenomenon observed is independent on functional J-protein-Hsp70 interactions. This technology has great potential for not only enhancing the production of recombinant proteins, but also to treat conformational diseases such as cystic fibrosis, and Alpha-1 antitrypsin deficiency.

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References

Sjobring U, Bjorck L, Kastern W . Streptococcal protein G. Gene structure and protein binding properties. J Biol Chem. 1991; 266(1):399-405. View

Sprague E, Martin W, Bjorkman P . pH dependence and stoichiometry of binding to the Fc region of IgG by the herpes simplex virus Fc receptor gE-gI. J Biol Chem. 2004; 279(14):14184-93. DOI: 10.1074/jbc.M313281200. View

Yu M, Lee K, Kim J . The Z type variation of human alpha 1-antitrypsin causes a protein folding defect. Nat Struct Biol. 1995; 2(5):363-7. DOI: 10.1038/nsb0595-363. View

Hebert D, Molinari M . In and out of the ER: protein folding, quality control, degradation, and related human diseases. Physiol Rev. 2007; 87(4):1377-408. DOI: 10.1152/physrev.00050.2006. View

Ellgaard L, Helenius A . Quality control in the endoplasmic reticulum. Nat Rev Mol Cell Biol. 2003; 4(3):181-91. DOI: 10.1038/nrm1052. View

Chen B, Retzlaff M, Roos T, Frydman J . Cellular strategies of protein quality control. Cold Spring Harb Perspect Biol. 2011; 3(8):a004374. PMC: 3140689. DOI: 10.1101/cshperspect.a004374. View

McClellan A, Tam S, Kaganovich D, Frydman J . Protein quality control: chaperones culling corrupt conformations. Nat Cell Biol. 2005; 7(8):736-41. DOI: 10.1038/ncb0805-736. View

Sprague E, Wang C, Baker D, Bjorkman P . Crystal structure of the HSV-1 Fc receptor bound to Fc reveals a mechanism for antibody bipolar bridging. PLoS Biol. 2006; 4(6):e148. PMC: 1450327. DOI: 10.1371/journal.pbio.0040148. View

Tabata Y, Chen W, Warrier M, Gibson A, Daines M, Khurana Hershey G . Allergy-driven alternative splicing of IL-13 receptor alpha2 yields distinct membrane and soluble forms. J Immunol. 2006; 177(11):7905-12. DOI: 10.4049/jimmunol.177.11.7905. View

10.

Gao C, Yam A, Wang X, Magdangal E, Salisbury C, Peretz D . Aβ40 oligomers identified as a potential biomarker for the diagnosis of Alzheimer's disease. PLoS One. 2011; 5(12):e15725. PMC: 3012719. DOI: 10.1371/journal.pone.0015725. View

11.

Genevaux P, Schwager F, Georgopoulos C, Kelley W . Scanning mutagenesis identifies amino acid residues essential for the in vivo activity of the Escherichia coli DnaJ (Hsp40) J-domain. Genetics. 2002; 162(3):1045-53. PMC: 1462316. DOI: 10.1093/genetics/162.3.1045. View

12.

DeLano W, Ultsch M, de Vos A, Wells J . Convergent solutions to binding at a protein-protein interface. Science. 2000; 287(5456):1279-83. DOI: 10.1126/science.287.5456.1279. View

13.

Papp E, Csermely P . Chemical chaperones: mechanisms of action and potential use. Handb Exp Pharmacol. 2006; (172):405-16. DOI: 10.1007/3-540-29717-0_16. View

14.

Bukau B, Weissman J, Horwich A . Molecular chaperones and protein quality control. Cell. 2006; 125(3):443-51. DOI: 10.1016/j.cell.2006.04.014. View

15.

Delic M, Gongrich R, Mattanovich D, Gasser B . Engineering of protein folding and secretion-strategies to overcome bottlenecks for efficient production of recombinant proteins. Antioxid Redox Signal. 2014; 21(3):414-37. DOI: 10.1089/ars.2014.5844. View

16.

Zhang J, Hilton D, Willson T, McFarlane C, Roberts B, Moritz R . Identification, purification, and characterization of a soluble interleukin (IL)-13-binding protein. Evidence that it is distinct from the cloned Il-13 receptor and Il-4 receptor alpha-chains. J Biol Chem. 1997; 272(14):9474-80. DOI: 10.1074/jbc.272.14.9474. View

17.

. American Thoracic Society/European Respiratory Society statement: standards for the diagnosis and management of individuals with alpha-1 antitrypsin deficiency. Am J Respir Crit Care Med. 2003; 168(7):818-900. DOI: 10.1164/rccm.168.7.818. View

18.

Lomas D, Evans D, Finch J, Carrell R . The mechanism of Z alpha 1-antitrypsin accumulation in the liver. Nature. 1992; 357(6379):605-7. DOI: 10.1038/357605a0. View

19.

Jeong Y, Kang H, Bae K, Kim M, Chung S . Efficient selection of IgG Fc domain-binding peptides fused to fluorescent protein using E. coli expression system and dot-blotting assay. Peptides. 2009; 31(2):202-6. DOI: 10.1016/j.peptides.2009.12.009. View

20.

Kraut-Cohen J, Gerst J . Addressing mRNAs to the ER: cis sequences act up!. Trends Biochem Sci. 2010; 35(8):459-69. DOI: 10.1016/j.tibs.2010.02.006. View