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Identification and characterization of a novel cartilage gene product CILP, which is an early indicator of osteoarthritis.

Lorenzo, Pilar LU (1998)
Abstract
The aim of this work was to study alterations in the matrix composition of human articular cartilage in different stages of osteoarthritis and to isolate and characterize one of the proteins that is altered in the osteoarthritic process. The metabolism of articular cartilage in different stages of osteoarthritis including early asymptomatic OA was monitored in a culture system of metabolically labeled cartilage explants. The labeled cartilage was extracted and the components separated at the analytical level in a two-dimensional system of ion exchange chromatography (Mono Q) in the first dimension, followed by an electrophoretic separation (SDS-polyacrylamide gel electrophoresis) of each effluent fractions. The labeled components were... (More)
The aim of this work was to study alterations in the matrix composition of human articular cartilage in different stages of osteoarthritis and to isolate and characterize one of the proteins that is altered in the osteoarthritic process. The metabolism of articular cartilage in different stages of osteoarthritis including early asymptomatic OA was monitored in a culture system of metabolically labeled cartilage explants. The labeled cartilage was extracted and the components separated at the analytical level in a two-dimensional system of ion exchange chromatography (Mono Q) in the first dimension, followed by an electrophoretic separation (SDS-polyacrylamide gel electrophoresis) of each effluent fractions. The labeled components were visualized by fluorography and quantitated by densitometric analysis. Several matrix components showed an altered synthesis and contents. One of those components, a 92 kDa protein, was further characterized after purification from human articular cartilage. The protein is a synthetic product of the chondrocytes and is restricted to cartilaginous tissues with a specific distribution in the middle layer of the normal articular cartilage. This specific distribution gave the name to the protein, CILP, (Cartilage Intermediate Layer Protein). Its deduced primary structure is quite unique, with very limited homology to other known proteins. In addition the gene encodes for another protein homologous to a porcine nucleotide pyrophosphohydrolase. The human CILP gene spans about 15.3 kbp of genomic DNA and it is organized in 9 exons interrupted by 8 introns. CILP is coded from exon 2 to exon 9, where the latter also codes for the entire human nucleotide pyrophosphohydrolase. Also particular to the gene was that one intron (intron 8) did not follow the "gt-ag" rule, but belongs to the minor classes of pre-mRNA introns containing "at-ac" at their 5’ and 3’ respectively. The gene was mapped to the chromosome 15q22, however no musculoskeletal diseases have been linked to this region. (Less)
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author
supervisor
opponent
  • Prof Vuorio, Eero, University of Turku, Kiinamyllyynkatu 10, SF-20520 Turku, Finland.
organization
publishing date
type
Thesis
publication status
published
subject
keywords
muscle system, Skeleton, matrix protein, NTPPHase, CILP, osteoarthritis, human cartilage, rheumatology locomotion, Skelett, muskelsystem, reumatologi
pages
50 pages
publisher
Department of Experimental Medical Science, Lund Univeristy
defense location
Kemicentrum, Hörsal A. Sölvegatan 39, Lund.
defense date
1998-12-01 09:00:00
external identifiers
  • other:ISRN: LUMEDW/MECM--98/1021-SE
ISBN
91-628-3232-8
language
English
LU publication?
yes
additional info
The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Department of Experimental Medical Science (013210000), Connective Tissue Biology (013230151)
id
044434a3-115b-4510-a0d6-b852015d8912 (old id 39088)
date added to LUP
2016-04-04 11:03:19
date last changed
2018-11-21 21:02:23
@phdthesis{044434a3-115b-4510-a0d6-b852015d8912,
  abstract     = {{The aim of this work was to study alterations in the matrix composition of human articular cartilage in different stages of osteoarthritis and to isolate and characterize one of the proteins that is altered in the osteoarthritic process. The metabolism of articular cartilage in different stages of osteoarthritis including early asymptomatic OA was monitored in a culture system of metabolically labeled cartilage explants. The labeled cartilage was extracted and the components separated at the analytical level in a two-dimensional system of ion exchange chromatography (Mono Q) in the first dimension, followed by an electrophoretic separation (SDS-polyacrylamide gel electrophoresis) of each effluent fractions. The labeled components were visualized by fluorography and quantitated by densitometric analysis. Several matrix components showed an altered synthesis and contents. One of those components, a 92 kDa protein, was further characterized after purification from human articular cartilage. The protein is a synthetic product of the chondrocytes and is restricted to cartilaginous tissues with a specific distribution in the middle layer of the normal articular cartilage. This specific distribution gave the name to the protein, CILP, (Cartilage Intermediate Layer Protein). Its deduced primary structure is quite unique, with very limited homology to other known proteins. In addition the gene encodes for another protein homologous to a porcine nucleotide pyrophosphohydrolase. The human CILP gene spans about 15.3 kbp of genomic DNA and it is organized in 9 exons interrupted by 8 introns. CILP is coded from exon 2 to exon 9, where the latter also codes for the entire human nucleotide pyrophosphohydrolase. Also particular to the gene was that one intron (intron 8) did not follow the "gt-ag" rule, but belongs to the minor classes of pre-mRNA introns containing "at-ac" at their 5’ and 3’ respectively. The gene was mapped to the chromosome 15q22, however no musculoskeletal diseases have been linked to this region.}},
  author       = {{Lorenzo, Pilar}},
  isbn         = {{91-628-3232-8}},
  keywords     = {{muscle system; Skeleton; matrix protein; NTPPHase; CILP; osteoarthritis; human cartilage; rheumatology locomotion; Skelett; muskelsystem; reumatologi}},
  language     = {{eng}},
  publisher    = {{Department of Experimental Medical Science, Lund Univeristy}},
  school       = {{Lund University}},
  title        = {{Identification and characterization of a novel cartilage gene product CILP, which is an early indicator of osteoarthritis.}},
  year         = {{1998}},
}