Chemical characterization of a lambda I amyloid protein isolated from formalin-fixed and paraffin-embedded tissue sections.
Amyloid protein was isolated from formalin-fixed paraffin-embedded heart tissue sections from a patient with primary (AL) amyloidosis by extraction with 6 M guanidine HCl. SDS-PAGE analysis of extracted material showed a major band at 16 kDa and a minor band at 18 kDa. Edman degradation analysis before and after pyroglutamate aminopeptidase treatment showed that the amyloid protein contained N-terminal pyroglutamic acid and was derived from an immunoglobulin lambda light chain. Analysis of tryptic peptides from the extract identified the amyloid protein as a lambda I. Of particular interest is that almost the entire amyloid protein amino acid sequence could be obtained from the cardiac sections. These results demonstrate that formalin-fixed paraffin-embedded tissue sections can be used for extensive biochemical characterization of amyloid proteins and will become a valuable source for isolation and extensive biochemical characterization of amyloid proteins as they are now a valuable source for isolation of DNA for genetic analysis.
Yazaki
Masahide
M
Department of Pathology and Laboratory Medicine, Indiana University School of Medicine, 635 Barnhill Drive, MS-128, Indianapolis, IN 46202-5126, USA.
Liepnieks
Juris J
JJ
Callaghan
John
J
Connolly
Charles E
CE
Benson
Merrill D
MD
D000595:Amino Acid Sequence; D000686:Amyloidosis / Q000150:complications / Q000235:genetics* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005557:Formaldehyde / Q000737:chemistry; D006333:Heart Failure / Q000209:etiology / Q000378:metabolism*; D006801:Humans; D007146:Immunoglobulin lambda-Chains / Q000032:analysis / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D007150:Immunohistochemistry; D008875:Middle Aged; D009206:Myocardium / Q000737:chemistry / Q000378:metabolism* / Q000473:pathology; D010265:Paraproteinemias / Q000150:complications / Q000235:genetics* / Q000378:metabolism; D017421:Sequence Analysis
D007146:Immunoglobulin lambda-Chains; D005557:Formaldehyde