Length variation in the keratan sulfate domain of mammalian aggrecan

Abstract

The keratan sulfate domain of aggrecan consists of a series of tandemly repeating hexapeptides which have the consensus sequence Glu-Glu/Lys-Pro-Phe-Pro-Ser, where the serine side-chains presumably provide sites for the attachment of keratan sulfate (KS) chains. The number of hexapeptide repeats varies between species, ranging from four in rat (Doege et al., 1987) and mouse (Walcz et al., 1992) to 13 in human (Doege et al., 1991) and 23 in bovine aggrecan (Antonsson et al., 1989). Chicken aggrecan (Chandrasekaran and Tanzer, 1992) does not contain a KS domain with a recognizable hexapeptide motif. The extent of this variation among mammalian and avian species is not known, and there is currently no explanation to predict how differences in the size of the KS domain would affect aggrecan function. We used polymerase chain reaction (PCR) to amplify the portion of the human, canine and porcine aggrecan gene that codes for the KS domain. We sequenced the amplified products in each case. Human aggrecan, with 13 hexapeptide repeats (Doege et al., 1987), was used as reference and found to be essentially identical to published data. The canine and porcine KS domains consisted of six and ten hexapeptide repeats respectively. The same PCR protocol was used to amplify the KS domain from genomic DNA of eight other mammalian species. Comparison of the size of these amplified products, as determined by agarose gel electrophoresis, with those for which sequence data are available allowed us to estimate the number of repeats in the KS domain. In almost half the species examined, the KS domain consisted of 13 hexapeptide repeats.