Detection and investigation of the molecular nature of low-molecular-mass copper ions in isolated rheumatoid knee-joint synovial fluid.
Low-molecular-mass copper(II) species have been detected and quantified in ultrafiltrates (n = 7) of rheumatoid synovial fluid (SF) by a highly-sensitive HPLC-based assay system with the ability to determine Cu(II) concentrations of < 10(-7) mol.dm-3. High field 1H NMR spectroscopy demonstrated that addition of Cu(II)(aq.) to isolated samples of RA SF ultrafiltrates resulted in complexation by histidine > alanine > formate > threonine > lactate > tyrosine > phenylalanine, their effectiveness in this context being in the given order. CD spectra of Cu(II)-treated samples of intact SF exhibited absorption bands typical of copper(II)-albumin complexes, in addition to a band attributable to a low-molecular-mass histidinate complex (lambda min 610 nm). Since both albumin and histidine are potent radical scavengers, these results indicate that any .OH radical generated from bound copper ions will be 'site-specifically' scavenged. Hence, low-molecular-mass copper complexes with the ability to promote the generation of .OH radical which can then escape from the metal ion co-ordination sphere (and in turn, cause damage to critical biomolecules) appear to be absent from inflammatory SF.
Naughton
D P
DP
Inflammation Research Group, London Hospital Medical College, UK.
Knappitt
J
J
Fairburn
K
K
Gaffney
K
K
Blake
D R
DR
Grootveld
M
M
D000328:Adult; D000596:Amino Acids / Q000032:analysis* / Q000302:isolation & purification; D001172:Arthritis, Rheumatoid / Q000097:blood / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002942:Circular Dichroism; D003300:Copper / Q000032:analysis* / Q000097:blood / Q000302:isolation & purification; D016166:Free Radical Scavengers; D006801:Humans; D017665:Hydroxyl Radical; D009682:Magnetic Resonance Spectroscopy; D008875:Middle Aged; D012016:Reference Values; D012680:Sensitivity and Specificity; D013582:Synovial Fluid / Q000737:chemistry*; D014462:Ultrafiltration
D000596:Amino Acids; D016166:Free Radical Scavengers; D017665:Hydroxyl Radical; D003300:Copper