Cloning of rat 92‐kDa type IV collagenase and expression of an active recombinant catalytic domain

Xia, Ying; García, Georgia Earnest; Chen, Shizhong; Wilson, Curtis B.; Li, Feng

doi:10.1016/0014-5793(96)00185-8

Cited by 21 publications

(18 citation statements)

References 30 publications

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“…The coding region was subcloned in an expression vector (pETM1) (37) to express a His-tagged recombinant mouse OB protein. The recombinant OB was expressed, purified, and refolded following a previously described procedure (38). Polyclonal antibodies were raised by immunizing a rabbit with the recombinant mouse OB and tested for biological activity to reduce body weight in ob/ob and control mice as described elsewhere.…”

Section: Methodsmentioning

confidence: 99%

Corticotropin-releasing Factor and Adrenocorticotrophic Hormone as Potential Central Mediators of OB Effects

Raber¹,

Chen²,

Mucke³

et al. 1997

Journal of Biological Chemistry

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OB (leptin) has been identified as a factor that suppresses appetite and stimulates metabolism. Attention has focused on the hypothalamus as its potential site of action, but OB could also act on other brain regions. In addition, the paradox of high OB levels in obese humans remains unresolved. Here we show in mice that both the long and short form of the OB receptor are expressed not only in the hypothalamus but also in the amygdala and pituitary. Recombinant murine OB elicited the release of corticotropin-releasing factor from superfused brain slice preparations containing hypothalamus or amygdala. Because corticotropin-releasing factor inhibits appetite and stimulates metabolism, it may be a key mediator of central OB effects. Recombinant OB also induced pituitary release of adrenocorticotrophic hormone. Because adrenocorticotrophic hormone-induced elevation of plasma glucocorticoid levels can inhibit corticotropin-releasing factor release via negative feedback, the OB effects on pituitary adrenocorticotrophic hormone release may be pertinent to human obesity, which combines increased plasma glucocorticoid levels with elevated levels of OB. An imbalance between the effects of OB on corticotropin-releasing factor release from the hypothalamus and on adrenocorticotrophic hormone release from the pituitary could contribute to obesity.Obesity is a cause of serious health problems, and new research on the ob gene, which encodes an adipose tissue-derived hormone (OB or leptin) that suppresses appetite and stimulates metabolism (1, 2), raises hopes for therapeutical intervention. Little is known about the molecular factors mediating OB effects. Increasing evidence supports a central site of action for OB. Infusion of OB into the lateral ventricles of normal or OB-deficient ob/ob mice decreases feeding (3). Although the hypothalamus has been identified as a potential site of action of OB (4), OB may also act on other regions. Short and long forms of the OB receptor (OB-R) 1 differing in the length of their intracellular domain are formed by differential splicing, but only the long OB-R form can activate janus kinase (JAK) and lead to phosphorylation of signal transducers and activators of transcription (STAT) proteins. The JAK/STAT pathway has been proposed to mediate the effects of OB on body weight (5). The function of the short OB-R form has not been identified yet, but this form could also have an important role in mediating central effects of OB.Although neuropeptide Y has been implicated as a potential central mediator of OB effects (4), recent evidence indicates that additional factors may be involved (6). OB-deficient ob/ob mice treated with OB show strong Fos protein immunoreactivity in the paraventricular nucleus (PVN) (7). It is interesting in this context that corticotropin-releasing factor (CRF) acting at the PVN inhibits food intake and stimulates metabolic rate in genetically obese animals and lean controls (8 -10). Furthermore, conditions that increase hypothalamic CRF production also diminish food in...

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Section: Methodsmentioning

confidence: 99%

Corticotropin-releasing Factor and Adrenocorticotrophic Hormone as Potential Central Mediators of OB Effects

Raber¹,

Chen²,

Mucke³

et al. 1997

Journal of Biological Chemistry

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“…RNase protection assays were performed as described 30 using a kit (Torrey Pines Biologicals, Houston, TX) with corresponding probes labeled with [ 32 P]UTP. Total RNA from organs and cells was isolated using Trizol (Gibco, BRL, Grand Island, NY).…”

Section: Rnase Protection Assaymentioning

confidence: 99%

“…30 The cDNA sequence encoding the human Slit2 protein from amino acids 1-300 (GenBank Accession No. XM 039647) was generated by PCR.…”

Section: Production Of Rabbit Polyclonal Anti-human Slit2 Antiseramentioning

confidence: 99%

Modulation of Inflammation by Slit Protein In Vivo in Experimental Crescentic Glomerulonephritis

Kanellis

García

et al. 2004

The American Journal of Pathology

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A basic conservation of cell migration guidance mechanisms in the nervous and immune systems was proposed when Slit, known for its role in axon guidance, was found to inhibit chemokine-induced leukocyte chemotaxis in vitro. These studies examined the role of Slit2 in modulating inflammation in vivo. In a rat model of glomerulonephritis, endogenous glomerular Slit2 expression fell after disease induction, and its inhibition during the early disease period accelerated inflammation. Ex vivo glomerular leukocytes showed decreased chemokine and chemoattractantinduced chemotaxis in response to Slit2, suggesting an anti-inflammatory role for glomerular Slit2. In contrast to the effect of inhibition, glomerulonephritis was ameliorated by systemic Slit2 administration. Slit2 treatment improved disease histologically and also improved renal function when given early in the disease course. Leukocytes harvested from rats receiving Slit2 showed decreased monocyte chemoattractant protein-1 (MCP)-1-mediated migration, consistent with a peripheral Slit2 effect. In keeping with this functional alteration, Slit2-mediated inhibition of RAW264.7 cell chemotaxis was associated with decreased levels of active cdc42 and Rac1, implicating GTPases in leukocyte Slit2 signaling.

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“…The MMP-9 promoter has several transcription factor binding motifs, as shown schematically in Figure 1, including three TPA response elements or AP-1 sites (TRE-1, -2 and -3), an NFkB site, an Ets site, an SP-1 site, and a GT box or retinoblastoma binding element (RBE) (Fini et al, 1994;Huhtala et al, 1990;Masure et al, 1993;Xia et al, 1996). Also, there is a 42 nucleotide CA (purine-pyrimidine) repeat whose function in the promoter is unknown.…”

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confidence: 99%

Transcriptional activation of the matrix metalloproteinase-9 gene in an H-ras and v-myc transformed rat embryo cell line

et al. 1997

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The 92 kd type IV collagenase/gelatinase (MMP-9) is important in mediating basement membrane and extracellular matrix degradation in metastasis. Because MMP-9 is made in tumor cells, but not in quiescent normal cells, we wished to identify the transcriptional elements responsible for its synthesis in tumor cells. We chose to characterize transcriptional regulation of the MMP-9 gene in a highly metastatic H-ras and v-myc transformed rat embryo cell line which overexpresses MMP-9. Using transient transfection of reporter gene constructs containing either 5'-deleted or mutated MMP-9 promoter fragments, as well as electrophoretic mobility shift assays, we have demonstrated that multiple transcription factor consensus binding motifs in the promoter, including those for NFkB, SP-1, Ets, AP-1, and a retinoblastoma binding element, participate in transcriptional regulation of MMP-9 expression in this cell line. Also, deletion of an alternating purinepyrimidine tract in the downstream promoter was found to decrease transcriptional activity, suggesting that promoter conformation may be important in MMP-9 regulation. Thus multiple pathways leading to activation of NFkB, SP-1, Ets, AP-1, and retinoblastoma binding factors in tumor cells all may contribute to MMP-9 transcription and hence to metastasis.

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Cloning of rat 92‐kDa type IV collagenase and expression of an active recombinant catalytic domain

Cited by 21 publications

References 30 publications

Corticotropin-releasing Factor and Adrenocorticotrophic Hormone as Potential Central Mediators of OB Effects

Corticotropin-releasing Factor and Adrenocorticotrophic Hormone as Potential Central Mediators of OB Effects

Modulation of Inflammation by Slit Protein In Vivo in Experimental Crescentic Glomerulonephritis

Transcriptional activation of the matrix metalloproteinase-9 gene in an H-ras and v-myc transformed rat embryo cell line

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