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J. Biol. Chem., Vol. 277, Issue 4, 2657-2665, January 25, 2002
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,
From the Sunnybrook and Women's College Health Sciences Centre and
Department of Laboratory Medicine and Pathobiology, University of
Toronto, Toronto M4N 3M5, Canada
Aggrecan is the major proteoglycan in the
extracellular matrix of cartilage. A notable exception is nanomelic
cartilage, which lacks aggrecan in its matrix. The example of nanomelia
and other evidence leads us to believe that the G3 domain plays an
important role in aggrecan processing, and it has indeed been confirmed that G3 allows glycosaminoglycan (GAG) chain attachment and product secretion. However, it is not clear how G3, which contains at least a
carbohydrate recognition domain (CRD) and a complement binding protein
(CBP) motif, plays these two functional roles. The present study was
designed to dissect the mechanisms of this phenomenon and specially 1)
to determine the effects of various cysteine residues in GAG
modification and product secretion as well as 2) to investigate which
of the two processing events is the critical step in the product
processing. Our studies demonstrated that removal of the two
amino-terminal cysteines in the CRD motif and the single cysteine in
the amino terminus of CBP inhibited secretion of CRD and CBP. Use of
the double mutant CRD construct also allowed us to observe a deviation
from the usual strict coupling of GAG modification and product
secretion steps. The presence of a small chondroitin sulfate fragment
overcame the secretion-inhibitory effects once the small chondroitin
sulfate fragment was modified by GAG.
This author, deceased February 22, 2001, was on sabbatical from the
University of Calgary.
§
A New Investigator of Canadian Institutes of Health Research and
recipient of a Premier's Research Excellence Award from the Province
of Ontario. To whom correspondence should be addressed: Research Bldg.,
Sunnybrook and Women's College Health Sciences Centre, 2075 Bayview
Ave., Toronto M4N 3M5, Canada. Tel.: 416-480-5874; Fax: 416-480-5737;
E-mail: Burton.Yang@swchsc.on.ca.
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