Post-translational aging of proteins in osteoarthritic cartilage and synovial fluid as measured by isomerized aspartate.
BACKGROUND
Aging proteins undergo non-enzymatic post-translational modification, including isomerization and racemization. We hypothesized that cartilage with many long-lived components could accumulate non-enzymatically modified amino acids in the form of isomerized aspartate and that its liberation due to osteoarthritis (OA)-related cartilage degradation could reflect OA severity.


METHODS
Articular cartilage and synovial fluid were obtained from 14 randomly selected total knee arthroplasty cases (56 to 79 years old) and non-arthritis cartilage from 8 trauma cases (51 to 83 years old). Paired lesional cartilage and non-lesioned OA cartilage were graded histologically using a modified Mankin system. Paired cartilage and synovial fluids were assayed for isomerized aspartate, phosphate-buffered saline/EDTA (ethylenediaminetetraacetic acid) extractable glycosaminoglycans, and total protein. Macroscopically normal non-lesioned OA cartilage was separated into superficial and deep regions when cartilage thickness was at least 3 mm (n = 6).


RESULTS
Normalized to cartilage wet weight, normal cartilage and deep non-lesioned OA cartilage contained significantly (P < 0.05) more isomerized aspartate than superficial non-lesioned OA cartilage and lesioned cartilage. Synovial fluid isomerized aspartate correlated positively (R2 = 0.53, P = 0.02) and glycosaminoglycans correlated negatively (R2 = 0.42, P = 0.04) with histological OA lesion severity. Neither synovial fluid isomerized aspartate nor glycosaminoglycans nor total protein correlated with histological scores of non-lesioned areas.


CONCLUSIONS
We show for the first time that human cartilage and synovial fluid contain measurable quantities of an isomerized amino acid and that synovial fluid concentrations of isomerized aspartate reflected severity of histological OA. Further assessment is warranted to identify the cartilage proteins containing this modification and to assess the functional consequences and biomarker applications of this analyte in OA.
Department of Medicine, Duke University, Durham, NC 27710, USA. Jonathan.Catterall@duke.edu
Jonathan B
JB
Catterall
Daniel
D
Barr
Michael
M
Bolognesi
Robert D
RD
Zura
Virginia B
VB
Kraus
D000368:Aged; D000369:Aged, 80 and over; D001224:Aspartic Acid / Q000737:chemistry / Q000378:metabolism*; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D002358:Cartilage, Articular / Q000378:metabolism* / Q000473:pathology; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D007536:Isomerism; D008875:Middle Aged; D020370:Osteoarthritis, Knee / Q000378:metabolism* / Q000473:pathology; D011499:Protein Processing, Post-Translational / Q000502:physiology*; D011506:Proteins / Q000378:metabolism*; D013582:Synovial Fluid / Q000378:metabolism*
D015415:Biomarkers; D006025:Glycosaminoglycans; D011506:Proteins; D001224:Aspartic Acid