Biochemical characterization of serum and urinary beta 2 microglobulin in end-stage renal disease patients.
Since the identification of beta 2 microglobulin (beta 2-M) in haemodialysis-associated amyloidosis, the biochemical characterization of the different forms of beta 2-M has been sought by several groups. New beta 2-M isoforms (pI 5.1 and lower) have been identified in amyloid deposits, and it has been suggested that they are of pathogenetic importance. The finding of N-terminal proteolysed beta 2-M in amyloid deposits prompted the hypothesis that proteolysis would render beta 2-M more amyloidogenic. Finally, a 'novel beta 2-M' (pI 5.2) with a single amino acid replacement (Asn by Asp at position 17) has been reported as possibly specific for patients with dialysis associated amyloidosis, and consequently proposed as 'the amyloidogenic' form. We purified beta 2-M from serum of a newly haemodialysed patient and from urine of a transplanted patient in the early recovery period. Both patients were clinically amyloid free. Three pure isoforms were obtained from serum (pI 5.7, 5.3, and 5.1) and only two from urine (5.7 and 5.3). Further purification of each isoform was obtained by HPLC in a C4 column. Sequence analysis showed that all isoforms had an intact N-terminus. Tryptic digestion of the serum isoforms was performed after alkylation with iodoacetic acid and the peptides were isolated by HPLC in a C18 column. The 5.3 and 5.1 isoforms had identical peptide patterns with the appearance of an early peak missing in the 5.7 form. The sequence of this peptide showed a replacement of the D 42 (Asp 42) by N (Asn) after K41 (Lys 41).(ABSTRACT TRUNCATED AT 250 WORDS)
Argilés
A
A
Centre de Recherche Biochimie Macromoleculaire (CRBM), CNRS 4PR 9008 INSERM Unit 249, Montpellier, France.
Derancourt
J
J
Jauregui-Adell
J
J
Mion
C
C
Demaille
J G
JG
D000328:Adult; D000595:Amino Acid Sequence; D000686:Amyloidosis / Q000209:etiology; D006801:Humans; D007676:Kidney Failure, Chronic / Q000097:blood / Q000378:metabolism* / Q000652:urine; D008297:Male; D008969:Molecular Sequence Data; D006435:Renal Dialysis / Q000009:adverse effects; D001613:beta 2-Microglobulin / Q000032:analysis* / Q000652:urine
D001613:beta 2-Microglobulin