Fractionation of Primary Amyloid Fibrils Characterization and Chemical Interaction of the Subunits

Jane B. Lian, Martha Skinner, Merrill Benson, Alan S. Cohen

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Amyloid fibrils of kappa origin from a patient with primary amyloidosis are dissociated in various denaturants and fractionated into their subunit components on Sepharose 6B. Solubilization of the fibrils in 4 M guanidine·HC1 followed by reduction and alkylation produced 22 000 and 17 000 dalton fractions. Without prior reduction and alkylation, these fractions exist as a high molecular weight protein which can be separated on Sepharose 6B. A high molecular weight protein can be directly dissociated from the amyloid fibril with 1% sodium dodecyl sulfate or 1 M NaCl. Reduction and alkylation of this material produces the two lower molecular weight fractions, i.e., 22 000 and 17 000. These have in the first 20 residues identical N-terminal amino acid sequences; they share immunologic identity and have similar tryptic peptide map profiles. Amino acid analysis of the 22 000 dalton fraction is identical with the intact immunoglobulin light chain isolated from the patient's serum. These data suggest that the insoluble amyloid fibril is the result of aggregation by disulfide linkages between the 22 000 and 17 000 dalton fractions.

Original languageEnglish (US)
Pages (from-to)167-176
Number of pages10
JournalBBA - Protein Structure
Volume491
Issue number1
DOIs
StatePublished - Mar 28 1977
Externally publishedYes

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Alkylation
Fractionation
Amyloid
Molecular Weight
Molecular weight
Sepharose
Immunoglobulin Light Chains
Amino Acids
Sodium Dodecyl Sulfate
Disulfides
Amino Acid Sequence
Proteins
Agglomeration
Peptides
Serum

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Fractionation of Primary Amyloid Fibrils Characterization and Chemical Interaction of the Subunits. / Lian, Jane B.; Skinner, Martha; Benson, Merrill; Cohen, Alan S.

In: BBA - Protein Structure, Vol. 491, No. 1, 28.03.1977, p. 167-176.

Research output: Contribution to journalArticle

Lian, Jane B. ; Skinner, Martha ; Benson, Merrill ; Cohen, Alan S. / Fractionation of Primary Amyloid Fibrils Characterization and Chemical Interaction of the Subunits. In: BBA - Protein Structure. 1977 ; Vol. 491, No. 1. pp. 167-176.
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