Cell-free synthesis of cartilage proteins: partial identification of proteoglycan core and link proteins

Treadwell, Benjamin V.; Mankin, David P.; Ho, Paul K.; Mankin, Henry J.

doi:10.1021/bi00551a043

Cited by 38 publications

(9 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The resulting minimum Mr for the unglycosylated protein core is 225000. This value is significantly lower than the estimates of 340000 for core protein translated in cell-free systems (Upholt et al, 1979;Treadwell et al, 1980). A more satisfactory agreement would be obtained when an a-helical conformation (Mr/L = 750 nm-1) is assumed for the extended domains.…”

Section: Discussionmentioning

confidence: 67%

Extended and globular protein domains in cartilage proteoglycans

Paulsson¹,

Mörgelin²,

Wiedemann³

et al. 1987

Biochemical Journal

137

102

View full text Add to dashboard Cite

Electron microscopy after rotary shadowing and negative staining of the large chondroitin sulphate proteoglycan from rat chondrosarcoma, bovine nasal cartilage and pig laryngeal cartilage demonstrated a unique multidomain structure for the protein core. A main characteristic is a pair of globular domains (diameter 6-8 nm), one of which forms the N-terminal hyaluronate-binding region. They are connected by a 25 nm-long rod-like domain of limited flexibility. This segment is continued by a 280 nm-long polypeptide strand containing most chondroitin sulphate chains (average length 40 nm) in a brush-like array and is terminated by a small C-terminal globular domain. The core protein showed a variable extent of degradation, including the loss of the C-terminal globular domain and sections of variable length of the chondroitin sulphate-bearing strand. The high abundance (30-50%) of the C-terminal domain in some extracted proteoglycan preparations indicated that this structure is present in the cartilage matrix rather than being a precursor-specific segment. It may contain the hepatolectin-like segment deduced from cDNA sequences corresponding to the 3'-end of protein core mRNA [Doege, Fernandez, Hassell, Sasaki & Yamada (1986) J. Biol. Chem. 261, 8108-8111; Sai, Tanaka, Kosher & Tanzer (1986) Proc. Natl. Acad. Sci. 83, 5081-5085; Oldberg, Antonsson & Heinegård (1987) Biochem. J. 243, 255-259].

show abstract

Section: Discussionmentioning

confidence: 67%

Extended and globular protein domains in cartilage proteoglycans

Paulsson¹,

Mörgelin²,

Wiedemann³

et al. 1987

Biochemical Journal

137

102

View full text Add to dashboard Cite

show abstract

“…The link proteins from bovine nasal cartilage are generally reported as two components with molecular weights of about 44,000 and 48,000 (1,40), although a smaller componcnt has also been observed (2). The link proteins isolated from bovine articular cartilage appear to be identical to those from bovine nasal cartilage (42). Link proteins also exist as two major components in both chicken and rabbit cartilage (1 1,24).…”

mentioning

confidence: 95%

Link protein shows species variation in its susceptibility to proteolysis

Liu

Cassidy

Allan

et al. 1992

Journal Orthopaedic Research

View full text Add to dashboard Cite

Human cartilage link protein exists as three native components, while equine, bovine, and porcine cartilage link protein exist as two and Swarm rat chondrosarcoma link protein exists as only one component. These nonhuman link protein components represent intact protein structures, and there is little evidence for proteolytically modified forms in nonhuman tissues. In human cartilage, the proteolytic production of modified link proteins increases with age, whereas high amounts of such products were not seen in the nonhuman tissues. However, the small amounts of link protein fragments that were observed in the nonhuman cartilages were of a similar size to their human counterparts. On digestion of human proteoglycan aggregate with stromelysin, rapid modification of the link protein components occurred, whereas the aggregates from nonhuman cartilages showed incomplete cleavage of their link protein components. The relative resistance of nonhuman link protein to stromelysin may in part be due to a unique amino acid substitution present near the enzymic cleave site.

show abstract

“…The aggregating proteoglycan found abundantly in cartilage is of particularly high molecular weight (Mr 1 x 106-4 x 10 6) (22). It consists of a protein core which, when newly synthesized, has a molecular weight of 300,000-350,000 (34,40,68,70,74) and to which are attached many chondroitin sulfate and keratan sulfate chains, and both o-linked and N-linked oligosaccharides (16,21,23). The protein core contains three main regions: a folded globular binding region, which forms a specific site for interaction with hyaluronate; an adjacent region rich in keratan…”

mentioning

confidence: 99%

Proteoglycan biosynthesis in chondrocytes: protein A-gold localization of proteoglycan protein core and chondroitin sulfate within Golgi subcompartments.

Ratcliffe¹,

Fryer²,

Hardingham³

1985

The Journal of Cell Biology

View full text Add to dashboard Cite

The intracellular pathway of cartilage proteoglycan biosynthesis was investigated in isolated chondrocytes using a protein A-gold electron microscopy immunolocalization procedure. Proteoglycans contain a protein core to which chondroitin sulfate and keratan sulfate chains and oligosaccharides are added in posttranslational processing. Specific antibodies have been used in this study to determine separately the distribution of the protein core and chondroitin sulfate components. In normal chondrocytes, proteoglycan protein core was readily localized only in smooth-membraned vesicles which co-labeled with ricin, indicating them to be galactose-rich medial/trans-Golgi cisternae, whereas there was only a low level of labeling in the rough endoplasmic reticulum. Chondroitin sulfate was also localized in medial/trans-Golgi cisternae of control chondrocytes but was not detected in other cellular compartments. In cells treated with monensin (up to 1.0/~M), which strongly inhibits proteoglycan secretion (Burditt, L. J., A. Ratcliffe, P. R. Fryer, and T. Hardingham, 1985, Biochim. Biophys. Acta., 844:247-255), there was greatly increased intracellular localization of proteoglycan protein core in both ricin-positive vesicles, and in ricin-negative vesicles (derived from cis-Golgi stacks) and in the distended rough endoplasmic reticulum. Chondroitin sulfate also increased in abundance after monensin treatment, but continued to be localized only in ricinpositive vesicles. The results suggested that the synthesis of chondroitin sulfate on proteoglycan only occurs in medial/trans-Golgi cisternae as a late event in proteoglycan biosynthesis. This also suggests that glycosaminoglycan synthesis on proteoglycans takes place in a compartment in common with events in the biosynthesis of both o-linked and N-linked oligosaccharides on other secretory glycoproteins.Proteoglycans are a specialized group of glycoproteins containing glycosaminoglycan chains. The aggregating proteoglycan found abundantly in cartilage is of particularly high molecular weight (Mr 1 x 106-4 x 10 6) (22). It consists of a protein core which, when newly synthesized, has a molecular weight of 300, 000-350,000 (34, 40, 68, 70, 74) and to which are attached many chondroitin sulfate and keratan sulfate chains, and both o-linked and N-linked oligosaccharides (16,21,23). The protein core contains three main regions: a folded globular binding region, which forms a specific site for interaction with hyaluronate; an adjacent region rich in keratan

show abstract

Cell-free synthesis of cartilage proteins: partial identification of proteoglycan core and link proteins

Cited by 38 publications

References 16 publications

Extended and globular protein domains in cartilage proteoglycans

Extended and globular protein domains in cartilage proteoglycans

Link protein shows species variation in its susceptibility to proteolysis

Proteoglycan biosynthesis in chondrocytes: protein A-gold localization of proteoglycan protein core and chondroitin sulfate within Golgi subcompartments.

Contact Info

Product

Resources

About