TRAMP (Tyrosine Rich Acidic Matrix Protein), a Protein that Co-purifies with Lysyl Oxidase from Porcine Skin

Cronshaw, Andrew D.; Macbeath, Jonathan R.E.; Shackleton, D. R.; Collins, John; Fothergill‐Gilmore, Linda A.; Hulmes, David J.S.

doi:10.1016/s0934-8832(11)80009-0

Cited by 37 publications

(17 citation statements)

References 36 publications

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“…Lane M shows the molecular weight markers (Bio-Rad). The 24-kDa protein that co-purifies with lysyl oxidase seen in lane P has previously been identified as the tyrosine-rich acidic matrix protein (TRAMP) (75,76 I, suramin increased lysyl oxidase activity in RS485 cell media by a factor of 2.2. This relatively small increase in enzyme activity in suramin-treated RS485 cells compared with mRNA increases is consistent with our finding of predominant production of unprocessed 50-kDa pro-lysyl oxidase summarized in Fig.…”

Section: Effect Of Suramin On Active Lysylmentioning

confidence: 97%

Autocrine Growth Factor Regulation of Lysyl Oxidase Expression in Transformed Fibroblasts

Palamakumbura

Sommer

Trackman

2003

Journal of Biological Chemistry

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Lysyl oxidase catalyzes oxidative deamination of peptidyl-lysine and hydroxylysine residues in collagens and lysine residues in elastin to form peptidyl aldehydes that are required for the formation of covalent crosslinks in normal extracellular matrix biosynthesis. Lysyl oxidase in addition has tumor suppressor activity, and phenotypic reversion of transformed cell lines is accompanied by increased lysyl oxidase expression. The mechanism of low expression of lysyl oxidase in tumor cells is unknown. The present study investigates the hypothesis that autocrine growth factor pathways maintain low lysyl oxidase expression levels in c-H-ras-transformed fibroblasts (RS485 cell line). Autocrine pathways were blocked with suramin, a general inhibitor of growth factor receptor binding, and resulted in more than a 10-fold increase in lysyl oxidase expression and proenzyme production. This regulation was found to be reversible and occurred at the transcriptional level determined using lysyl oxidase promoter/reporter gene assays. Function blocking anti-fibroblast growth factor-2 (FGF-2) antibody enhanced lysyl oxidase expression in the absence of suramin. Finally, the addition of FGF-2 to suramin-treated cells completely reversed suramin stimulation of lysyl oxidase mRNA levels. Data support that an FGF-2 autocrine pathway inhibits lysyl oxidase transcription in the tumorigenic-transformed RS485 cell line. This finding may be of therapeutic significance and, in addition, provides a new experimental approach to investigate the mechanism of the tumor suppressor activity of lysyl oxidase.

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Section: Effect Of Suramin On Active Lysylmentioning

confidence: 97%

Autocrine Growth Factor Regulation of Lysyl Oxidase Expression in Transformed Fibroblasts

Palamakumbura

Sommer

Trackman

2003

Journal of Biological Chemistry

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“…TRAMP was purified from the skins of stillborn piglets, by DEAE ion-exchange chromatography and selective interaction with Sephacryl S-400, followed by preparative reverse-phase chromatography on a Pharmacia ProRPC HR5/2 column, as described [14]. Polyclonal antiserum was raised against TRAMP by subcutaneous injection of New Zealand White rabbits with an initial dose of 200/.tg TRAMP emulsified in a 1 : 1 mixture of complete Freund's adjuvant, followed by a I00 /~g boost (in incomplete Freund's adjuvant) and then four further boosts injected every 6 weeks.…”

Section: Preparation Of Polyclonal Antiserummentioning

confidence: 99%

“…Briefly, 5 g tyrosine and 10 ml concentrated sulphuric acid (both pre-cooled to -20°C) were mixed and stirred rapidly on ice for 10 min, then neutralised with Ba(OH)2, and the sulphotyrosine separated from unreacted tyrosine on a Bio-Rad AG 50W-X8 column [22]. The elution position of the phenylthiocarbamyl (PTC) derivative of sulphotyrosine was determined on an Applied Biosystems 420A amino acid analyser [14] after derivatisation with phenylisothiocyanate. PTC-sulphotyrosine eluted between PTC-proline and phenylthiourea.…”

Section: Identification Of Sulphotyrosinementioning

confidence: 99%

“…Previous work has indicated that TRAMP is not a glycoprotein [15], and the effect of sulphatase treatment or mild acid hydrolysis on Alcian blue staining indicated that TRAMP contained sulphotyrosine [14]. To examine this possibility, immunoprecipitated TRAMP from the human fibroblast culture medium was separated by SDS-PAGE, then transferred to 435 ProBlott and subjected to alkaline hydrolysis.…”

Section: Eg Forbes Et Al Ifebs Letters 351 (1994) 433-436mentioning

confidence: 99%

“…During the course of purifying lysyl oxidase (32 kDa) from porcine skin, we recently characterised a protein (22 kDa) that co-purifies with the enzyme [14]. This new protein is acidic and rich in tyrosine, and we refer to it as TRAMP (tyrosine-rich acidic matrix protein).…”

Section: Introductionmentioning

confidence: 99%

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Tyrosine‐rich acidic matrix protein (TRAMP) is a tyrosine‐sulphated and widely distributed protein of the extracellular matrix

et al. 1994

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Tyrosine-rich acidic matrix protein (TRAMP; 22 kDa extracellular matrix protein; dermatopontin) is a protein that co-purifies with lysyl oxidase and with dermatan sulphate proteoglycans, with possible functions in cell-matrix interactions and matrix assembly. Using a rabbit polyclonal antiserum raised against porcine TRAMP, which cross-reacts with both the human and murine forms of the protein, we show by immunoblotting that TRAMP has a widespread tissue distribution, including skin, skeletal muscle, heart, lung, kidney, cartilage and bone. In cultures of human skin fibroblasts, TRAMP incorporates both [35S]sulphate and [3H]tyrosine and is secreted into the medium, as shown by immunoprecipitation. Amino acid analysis of immunoprecipitated TRAMP demonstrates that many of the tyrosine residues in TRAMP are sulphated.

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Reduced dermatopontin expression is a molecular link between uterine leiomyomas and keloids

Catherino

Leppert

Stenmark

et al. 2004

Genes Chromosomes & Cancer

152

148

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Uterine leiomyomas are prevalent estrogen-responsive clonal tumors, but the specific genetic alterations that contribute to their development have not been elucidated. To identify genes involved in the formation of leiomyomas, we used global expression profiling to compare clonal tumors with normal myometrium. Contrary to expectation, genes involved in estrogen action were not differentially expressed between leiomyoma and normal myometrium. Genes encoding extracellular-matrix proteins were prominently featured, suggesting their involvement in formation of a myofibroblast phenotype. Analysis of the extracellular matrix in the leiomyomas revealed a disordered collagen fibril orientation. Expression of the collagen-binding protein dermatopontin was found to be consistently decreased in leiomyoma by both reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time RT-PCR (mean underexpression = 9.41-fold) regardless of leiomyoma size, leiomyoma location, patient race, and patient age. This expression pattern was observed in 11 subjects and a total of 23 leiomyoma:myometrium pairs. Decreased expression of dermatopontin was also associated with keloid formation, a fibrotic disease that shares epidemiologic similarities with leiomyoma. Immunohistochemical studies of leiomyomas and keloids demonstrated reduced levels of dermatopontin in both tissues. In addition, ultrastructural analysis revealed that the orientation of the collagen fibrils in the keloid tissues strongly resembled that in the leiomyomas. Reduction in dermatopontin was associated with an increase in transforming growth factor-beta3 (TGFB3) mRNA levels in leiomyomas, whereas other genes involved in dermatopontin signaling were not differentially expressed. These findings suggest that leiomyoma development involves a myofibroblast cell phenotype characterized by dysregulation of genes encoding extracellular-matrix proteins. In particular, decreased expression of dermatopontin represents a molecular link between the leiomyoma and keloid phenotypes.

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TRAMP (Tyrosine Rich Acidic Matrix Protein), a Protein that Co-purifies with Lysyl Oxidase from Porcine Skin

Cited by 37 publications

References 36 publications

Autocrine Growth Factor Regulation of Lysyl Oxidase Expression in Transformed Fibroblasts

Autocrine Growth Factor Regulation of Lysyl Oxidase Expression in Transformed Fibroblasts

Tyrosine‐rich acidic matrix protein (TRAMP) is a tyrosine‐sulphated and widely distributed protein of the extracellular matrix

Reduced dermatopontin expression is a molecular link between uterine leiomyomas and keloids

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