scholarly journals The primary structure of human cartilage link protein

1990 ◽  
Vol 18 (5) ◽  
pp. 1292-1292 ◽  
Author(s):  
Jayesh Dudhia ◽  
Timothy E. Hardingham
Keyword(s):  
Primary Structure ◽  
Human Cartilage ◽  
Link Protein

scholarly journals Expression and Purification of Functionally Active Hyaluronan-binding Domains from Human Cartilage Link Protein, Aggrecan and Versican

2004 ◽  
Vol 280 (7) ◽  
pp. 5435-5448 ◽  
Author(s):  
Nicholas T. Seyfried ◽  
Gillian F. McVey ◽  
Andrew Almond ◽  
David J. Mahoney ◽  
Jayesh Dudhia ◽  
...  
Keyword(s):  
Expression And Purification ◽  
Human Cartilage ◽  
Link Protein ◽  
Binding Domains ◽  
Hyaluronan Binding

scholarly journals Variations in aggrecan structure modulate its susceptibility to aggrecanases

2003 ◽  
Vol 375 (1) ◽  
pp. 183-189 ◽  
Author(s):  
Peter J. ROUGHLEY ◽  
James BARNETT ◽  
Fengrong ZUO ◽  
John S. MORT
Keyword(s):  
Chondroitin Sulphate ◽  
Human Cartilage ◽  
In Vitro Degradation ◽  
Link Protein ◽  
Rich Domain ◽  
Sds Page ◽  
Lower Ability ◽  
Proteoglycan Aggregates ◽  
Fetal Bovine

Proteoglycan aggregates and purified aggrecan from adult and fetal bovine cartilage and adult and neonatal human cartilage were subjected to in vitro degradation by recombinant aggrecanase-1 and aggrecanase-2. The ability of the aggrecanases to cleave within the aggrecan IGD (interglobular domain) and CS2 domain (chondroitin sulphate-rich domain 2) was monitored by SDS/PAGE and immunoblotting. Aggrecanase-2 showed a similar ability to cleave within the IGD of adult and immature aggrecan, whereas aggrecanase-1 was less efficient in cleavage in the IGD of immature aggrecan, for both the bovine and the human substrates. Both aggrecanases showed a similar ability to cleave within the CS2 domain of bovine aggrecan irrespective of age, but showed a much lower ability to cleave within the CS2 domain of human aggrecan. Equivalent results were obtained whether aggrecan was present in isolation or as part of proteoglycan aggregates. When proteoglycan aggregates were used, neither aggrecanase was able to cleave link protein. Thus, for aggrecan cleavage by aggrecanases, variations in cleavage efficiency exist with respect to the species and age of the animal from which the aggrecan is derived and the type of aggrecanase being used.

scholarly journals Matrix metalloproteinases cleave at two distinct sites on human cartilage link protein

1993 ◽  
Vol 295 (2) ◽  
pp. 595-598 ◽  
Author(s):  
Q Nguyen ◽  
G Murphy ◽  
C E Hughes ◽  
J S Mort ◽  
P J Roughley
Keyword(s):  
Matrix Metalloproteinases ◽  
Degradation Products ◽  
Limited Proteolysis ◽  
Human Cartilage ◽  
Link Protein ◽  
Sequencing Studies ◽  
Proteoglycan Aggregates ◽  
Protein Components ◽  
Gelatinases A And B

The actions of human recombinant stromelysins-1 and -2, collagenase, gelatinases A and B and matrilysin on neonatal human proteoglycan aggregates were examined. With the exception of gelatinase B, aggrecan was degraded extensively by most metalloproteinases studied, whereas link protein showed only limited proteolysis. Sequencing studies of modified link protein components revealed that stromelysins-1 and -2, gelatinases A and B and collagenase cleaved specifically between His16 and Ile17, and matrilysin, stromelysin-2 and gelatinase A cleaved between Leu25 and Leu26. Cleavage at the former bond generated a link protein component with the same N-terminus as that isolated from newborn human cartilage. Based on previously determined in situ cleavage sites it is evident that matrix metalloproteinases are not solely responsible for the accumulation of link protein degradation products in adult human cartilage, indicating that additional proteolytic agents are involved in the normal catabolism of human cartilage matrix.

scholarly journals The origin of human cartilage proteoglycan link-protein heterogeneity and fragmentation during aging

1985 ◽  
Vol 232 (3) ◽  
pp. 805-812 ◽  
Author(s):  
J S Mort ◽  
B Caterson ◽  
A R Poole ◽  
P J Roughley
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Human Articular Cartilage ◽  
Human Cartilage ◽  
Link Protein ◽  
Cartilage Proteoglycan ◽  
Disulphide Bonds ◽  
Single Link ◽  
Chemical Deglycosylation

Human articular-cartilage link proteins are resolved into three components by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, indicative of three different structures. The action of the proteinase clostripain yields a single link-protein component with electrophoretic properties analogous to those of the smallest (most mobile) native link protein, suggesting that this link protein may be derived naturally from one or both of the larger molecules by proteolytic cleavage in situ. Upon chemical deglycosylation of native link protein two components are resolved, suggesting that two of the link proteins differ only in their degree and/or type of oligosaccharide substitution. This pattern is compatible with a proteolytic origin for the smallest link protein. During aging further proteolytic fragmentation occurs, though it is only apparent on reduction of disulphide bonds. This fragmentation occurs at identical sites in all three native link proteins, indicating the existence of a large region common to all the link proteins, which appears to consist predominantly of the C-terminal half of the molecules. These observations are compatible with the variation in oligosaccharide and proteolytic heterogeneity occurring at the N-terminus of the link proteins.

Isolation and sequence of cDNA clones for pig and human cartilage link protein

1989 ◽  
Vol 206 (4) ◽  
pp. 749-753 ◽  
Author(s):  
Jayesh Dudhia ◽  
Timothy E. Hardingham
Keyword(s):  
Cdna Clones ◽  
Human Cartilage ◽  
Link Protein

Complete amino acid sequence of human cartilage link protein (CRTL1) deduced from cDNA clones and chromosomal assignment of the gene

Genomics ◽  
1990 ◽  
Vol 8 (3) ◽  
pp. 562-567 ◽  
Author(s):  
Sherri L. Osborne-Lawrence ◽  
Andrea K. Sinclair ◽  
Robert C. Hicks ◽  
Stephen W. Lacey ◽  
Roger L. Eddy ◽  
...  
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Chromosomal Assignment ◽  
Cdna Clones ◽  
Human Cartilage ◽  
Link Protein ◽  
Complete Amino Acid Sequence

scholarly journals Human link protein gene: structure and transcription pattern in chondrocytes

1994 ◽  
Vol 303 (1) ◽  
pp. 329-333 ◽  
Author(s):  
J Dudhia ◽  
M T Bayliss ◽  
T E Hardingham
Keyword(s):  
Transcription Initiation ◽  
Protein Gene ◽  
Transcription Unit ◽  
Flanking Sequence ◽  
Human Cartilage ◽  
S1 Nuclease ◽  
Link Protein ◽  
Alternatively Spliced ◽  
Similar Distance ◽  
Age Range

We have examined the genomic organization and the transcription unit for the human link protein gene from genomic clones and RNA prepared from human cartilage over a wide age range. Five exons cover the gene which is greater than 60 kbp. Primer extension and S1 nuclease protection analysis revealed transcription initiation to be 315 bases upstream from the translation initiation codon in RNA derived from cartilage samples ranging from fetal to 53 years of age. The first exon size therefore is 289 bp and examination of the 5′ flanking sequence indicated a lack of a TATA box in close proximity to the transcription start, although a TATAA-like motif (TCTAA) was present at -75 bp. Such a sequence at a similar distance can serve as a promoter in the chicken link protein gene. The large first exon of 289 bp is similar to that of the chicken but contrasts with that described previously for human (96 bp) and rat (62 bp). We also analysed human link protein mRNA by PCR for the presence of an alternatively spliced exon that is present in rat mRNA in low abundance, but could not detect such transcripts. Equine and porcine mRNA contained this spliced form but the results suggested that this was expressed as a rare transcript.

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