Recombinant immunoglobulin variable domains generated from synthetic genes provide a system for in vitro characterization of light‐chain amyloid proteins

Priscilla Wilkins Stevens, Rosemarie Raffen, Deborah K. Hanson, Ya‐Li ‐L Deng, Maria Berrios‐Hammond, Florence A. Westholm, Marianne Schiffer, Fred J. Stevens, Charles Murphy, Alan Solomon, Manfred Eulitz, Ronald Wetzel

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Abstract

The primary structural features that render human monoclonal light chains amyloidogenic are presently unknown. To gain further insight into the physical and biochemical factors that result in the pathologic deposition of these proteins as amyloid fibrils, we have selected for detailed study three closely homologous protein products of the light‐chain variable‐region single‐gene family VkIV. Two of these proteins, REC and SMA, formed amyloid fibrils in vivo. The third protein, LEN, was excreted by the patient at levels of 50 g/day with no indication of amyloid deposits. Sequences of amyloidogenic proteins REC and SMA differed from the sequence of the nonpathogenic protein LEN at 14 and 8 amino acid positions, respectively, and these amino acid differences have been analyzed in terms of the three‐dimensional structure of the LEN dimer. To provide a replenishable source of these human proteins, we constructed synthetic genes coding for the REC, SMA, and LEN variable domains and expressed these genes in Escherichia coli. Immunochemical and biophysical comparisons demonstrated that the recombinant VkIV products have tertiary structural features comparable to those of the patient‐derived proteins. This well‐defined set of three clinically characterized human kIV light chains, together with the capability to produce these kIV proteins recombinantly, provide a system for biophysical and structural comparisons of two different amyloidogenic light‐chain proteins and a nonamyloidogenic protein of the same subgroup. This work lays the foundation for future investigations of the structural basis of light‐chain amyloidogenicity.

Original languageEnglish (US)
Pages (from-to)421-432
Number of pages12
JournalProtein Science
Volume4
Issue number3
DOIs
StatePublished - Jan 1 1995

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Synthetic Genes
Amyloidogenic Proteins
Immunoglobulins
Genes
Proteins
Amyloid
In Vitro Techniques
Immunoglobulin Domains
Light
Amino Acids
Amyloid Plaques
Dimers
Escherichia coli
Deposits

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Stevens, P. W., Raffen, R., Hanson, D. K., Deng, YL. L., Berrios‐Hammond, M., Westholm, F. A., ... Wetzel, R. (1995). Recombinant immunoglobulin variable domains generated from synthetic genes provide a system for in vitro characterization of light‐chain amyloid proteins. Protein Science, 4(3), 421-432. https://doi.org/10.1002/pro.5560040309

Recombinant immunoglobulin variable domains generated from synthetic genes provide a system for in vitro characterization of light‐chain amyloid proteins. / Stevens, Priscilla Wilkins; Raffen, Rosemarie; Hanson, Deborah K.; Deng, Ya‐Li ‐L; Berrios‐Hammond, Maria; Westholm, Florence A.; Schiffer, Marianne; Stevens, Fred J.; Murphy, Charles; Solomon, Alan; Eulitz, Manfred; Wetzel, Ronald.

In: Protein Science, Vol. 4, No. 3, 01.01.1995, p. 421-432.

Research output: Contribution to journalArticle

Stevens, PW, Raffen, R, Hanson, DK, Deng, YLL, Berrios‐Hammond, M, Westholm, FA, Schiffer, M, Stevens, FJ, Murphy, C, Solomon, A, Eulitz, M & Wetzel, R 1995, 'Recombinant immunoglobulin variable domains generated from synthetic genes provide a system for in vitro characterization of light‐chain amyloid proteins', Protein Science, vol. 4, no. 3, pp. 421-432. https://doi.org/10.1002/pro.5560040309
Stevens, Priscilla Wilkins ; Raffen, Rosemarie ; Hanson, Deborah K. ; Deng, Ya‐Li ‐L ; Berrios‐Hammond, Maria ; Westholm, Florence A. ; Schiffer, Marianne ; Stevens, Fred J. ; Murphy, Charles ; Solomon, Alan ; Eulitz, Manfred ; Wetzel, Ronald. / Recombinant immunoglobulin variable domains generated from synthetic genes provide a system for in vitro characterization of light‐chain amyloid proteins. In: Protein Science. 1995 ; Vol. 4, No. 3. pp. 421-432.
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