Expression and Purification of ADAMTSL5 in E. Coli and Validation of Activity in Psoriasis Serum

Overview

Journal Immun Inflamm Dis

Specialties Allergy & Immunology
Infectious Diseases

Date 2024 Dec 13

PMID 39670545

Authors

Ru Qin

Shangyang Li

Boheng Wu

Ruilan Lin

Yulin Yuan

Affiliations

Soon will be listed here.

Abstract

Purpose: A disintegrin and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5), a protein linked to psoriasis, was obtained by prokaryotic expression and purification for potential utilization as a new auxiliary diagnostic marker for psoriasis. It was subsequently applied in psoriasis research.

Experimental Design: The designed ADAMTSL5 gene was inserted into the pET-30a (+) vector and expressed in the BL21 (DE3) strain as a fusion protein. Following this, the recombinant ADAMTSL5 protein was purified using affinity chromatography. Purified ADAMTSL5, obtained in conjunction with magnetic beads, was directly employed in both psoriasis patients and healthy individuals.

Results: The results indicated that the molecular weight of the ADAMTSL5 protein obtained through prokaryotic expression and purification was approximately 27 kDa, with a protein concentration of 0.96 mg/mL. Analysis of anti-ADAMTSL5 levels in the serum of both psoriasis patients and healthy individuals using magnetic microparticle chemiluminescence demonstrated that anti-ADAMTSL5 levels in the serum of psoriasis patients surpassed those of healthy individuals, showing a significant difference with p < 0.001.

Conclusions: The experimental findings presented here may contribute to the utilization of ADAMTSL5 as a marker for diagnosing psoriasis, offering novel insights and experimental avenues for further investigation.

References

Dupire G, Droitcourt C, Hughes C, Le Cleach L . Antistreptococcal interventions for guttate and chronic plaque psoriasis. Cochrane Database Syst Rev. 2019; 3:CD011571. PMC: 6400423. DOI: 10.1002/14651858.CD011571.pub2. View

Schon M, Boehncke W . Psoriasis. N Engl J Med. 2005; 352(18):1899-912. DOI: 10.1056/NEJMra041320. View

Kim S, Bhonsle L, Besgen P, Nickel J, Backes A, Held K . Analysis of the paired TCR α- and β-chains of single human T cells. PLoS One. 2012; 7(5):e37338. PMC: 3359365. DOI: 10.1371/journal.pone.0037338. View

Diluvio L, Vollmer S, Besgen P, Ellwart J, Chimenti S, Prinz J . Identical TCR beta-chain rearrangements in streptococcal angina and skin lesions of patients with psoriasis vulgaris. J Immunol. 2006; 176(11):7104-11. DOI: 10.4049/jimmunol.176.11.7104. View

Arakawa A, Siewert K, Stohr J, Besgen P, Kim S, Ruhl G . Melanocyte antigen triggers autoimmunity in human psoriasis. J Exp Med. 2015; 212(13):2203-12. PMC: 4689169. DOI: 10.1084/jem.20151093. View

Yuan Y, Qiu J, Lin Z, Li W, Haley C, Mui U . Identification of Novel Autoantibodies Associated With Psoriatic Arthritis. Arthritis Rheumatol. 2019; 71(6):941-951. DOI: 10.1002/art.40830. View

Ramirez F, Sakai L . Biogenesis and function of fibrillin assemblies. Cell Tissue Res. 2009; 339(1):71-82. PMC: 2819175. DOI: 10.1007/s00441-009-0822-x. View

Banyai L, Patthy L . The NTR module: domains of netrins, secreted frizzled related proteins, and type I procollagen C-proteinase enhancer protein are homologous with tissue inhibitors of metalloproteases. Protein Sci. 1999; 8(8):1636-42. PMC: 2144412. DOI: 10.1110/ps.8.8.1636. View

Prinz J . Melanocytes: Target Cells of an HLA-C*06:02-Restricted Autoimmune Response in Psoriasis. J Invest Dermatol. 2017; 137(10):2053-2058. DOI: 10.1016/j.jid.2017.05.023. View

10.

Harden J, Krueger J, Bowcock A . The immunogenetics of Psoriasis: A comprehensive review. J Autoimmun. 2015; 64:66-73. PMC: 4628849. DOI: 10.1016/j.jaut.2015.07.008. View

11.

Muhr P, Zeitvogel J, Heitland I, Werfel T, Wittmann M . Expression of interleukin (IL)-1 family members upon stimulation with IL-17 differs in keratinocytes derived from patients with psoriasis and healthy donors. Br J Dermatol. 2011; 165(1):189-93. DOI: 10.1111/j.1365-2133.2011.10302.x. View

12.

Menssen A, Trommler P, Vollmer S, Schendel D, Albert E, Gurtler L . Evidence for an antigen-specific cellular immune response in skin lesions of patients with psoriasis vulgaris. J Immunol. 1995; 155(8):4078-83. View

13.

Apte S . A disintegrin-like and metalloprotease (reprolysin-type) with thrombospondin type 1 motif (ADAMTS) superfamily: functions and mechanisms. J Biol Chem. 2009; 284(46):31493-7. PMC: 2797218. DOI: 10.1074/jbc.R109.052340. View

14.

Favaro R, Facheris P, Formai A, Gargiulo L, Ibba L, Fiorillo G . Autoreactivity to self-antigens LL37 and ADAMTSL5 influences the clinical response to risankizumab in psoriatic patients. J Autoimmun. 2024; 147:103244. DOI: 10.1016/j.jaut.2024.103244. View

15.

Bader H, Wang L, Ho J, Tran T, Holden P, Fitzgerald J . A disintegrin-like and metalloprotease domain containing thrombospondin type 1 motif-like 5 (ADAMTSL5) is a novel fibrillin-1-, fibrillin-2-, and heparin-binding member of the ADAMTS superfamily containing a netrin-like module. Matrix Biol. 2012; 31(7-8):398-411. PMC: 3546522. DOI: 10.1016/j.matbio.2012.09.003. View

16.

Chang J, Smith L, Froning K, Schwabe B, Laxer J, Caralli L . CD8+ T cells in psoriatic lesions preferentially use T-cell receptor V beta 3 and/or V beta 13.1 genes. Proc Natl Acad Sci U S A. 1994; 91(20):9282-6. PMC: 44796. DOI: 10.1073/pnas.91.20.9282. View

17.

Vollmer S, Menssen A, Prinz J . Dominant lesional T cell receptor rearrangements persist in relapsing psoriasis but are absent from nonlesional skin: evidence for a stable antigen-specific pathogenic T cell response in psoriasis vulgaris. J Invest Dermatol. 2001; 117(5):1296-301. DOI: 10.1046/j.0022-202x.2001.01494.x. View

18.

Henseler T, Christophers E . Psoriasis of early and late onset: characterization of two types of psoriasis vulgaris. J Am Acad Dermatol. 1985; 13(3):450-6. DOI: 10.1016/s0190-9622(85)70188-0. View

19.

Gudjonsson J, Karason A, Antonsdottir A, Runarsdottir E, Hauksson V, Upmanyu R . Psoriasis patients who are homozygous for the HLA-Cw*0602 allele have a 2.5-fold increased risk of developing psoriasis compared with Cw6 heterozygotes. Br J Dermatol. 2003; 148(2):233-5. DOI: 10.1046/j.1365-2133.2003.05115.x. View

20.

Arechederra M, Bazai S, Abdouni A, Sequera C, Mead T, Richelme S . ADAMTSL5 is an epigenetically activated gene underlying tumorigenesis and drug resistance in hepatocellular carcinoma. J Hepatol. 2020; 74(4):893-906. DOI: 10.1016/j.jhep.2020.11.008. View