Cloning and expression of three rabbit kidney cDNAs encoding lauric acid .omega.-hydroxylases

Катруха, Г. С.; Silaev, A. B.; Katrukha, S. P.

doi:10.1021/bi00456a004

Cited by 62 publications

(25 citation statements)

References 23 publications

(45 reference statements)

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“…Lauric acid was therefore selected to avoid subsequent metabolism and to give a much simpler measure of CYP4A activity. Although it is reasonable to hypothesize that the individual rat CYP4A isoforms may have unique substrate specificities, similar to that described for the multiple rabbit CYP4A isoforms (32,33), there were no apparent differences in metabolism between these two substrates with WKY rat and SHR renal microsomes. Lauric acid -hydroxylation activity was consistent with arachidonic acid -hydroxylation, although the magnitude of the differences in activity between WKY rat and SHR kidneys was smaller with lauric acid.…”

Section: Cyp4a Expression and Activity In The Shr Kidney 369mentioning

confidence: 69%

Developmentally Regulated Expression of theCYP4AGenes in the Spontaneously Hypertensive Rat Kidney

et al. 1997

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SUMMARYThe CYP4A enzymes catalyze the formation of 20-hydroxyeicosatetraenoic acid (20-HETE), which has potent effects on the renal vasculature and tubular ion transport. Based on an increased 20-HETE formation in renal microsomes from spontaneously hypertensive rats, it has been proposed that increased expression of the CYP4A genes is an early event in the development of hypertension in these animals. To test this hypothesis, we developed RNase protection assays for specific detection of the individual CYP4A genes in the kidneys of spontaneously hypertensive and Wistar-Kyoto rats. Distinct age-dependent patterns of expression were observed for the individual CYP4A genes, with only CYP4A3 mRNA measurable in the kidneys of 1-week-old rats. CYP4A1 and CYP4A8 mRNA were detectable by 3 weeks of age and CYP4A2 mRNA at 5 weeks of age. The expression of CYP4A1 and CYP4A3 varied 4 -5-fold throughout development and was highest between 3 and 5 weeks of age, declining steadily thereafter to 20% of their maximal level by 9 weeks of age. CYP4A2 mRNA levels increased steadily between 5 and 9 weeks of age, whereas CYP4A8 mRNA levels were relatively constant throughout development. The CYP4A3 mRNA level was significantly increased 1.6 -2-fold in the cortex and outer medulla of 1-4-week-old spontaneously hypertensive rat kidneys relative to the corresponding level in the Wistar-Kyoto. A similar 1.4 -1.7-fold increase in CYP4A8 mRNA was also found in 3-and 4-weekold spontaneously hypertensive kidneys. Accompanying the increased expression of CYP4A3 and CYP4A8 mRNA in the prehypertensive rats were corresponding changes in functional CYP4A measured as either arachidonic acid or lauric acid -hydroxylase activity (1.4 -2.0-fold increases) and CYP4A protein levels. After 4 weeks of age, the level of CYP4A mRNA, enzyme activity, and protein were similar in the kidneys of Wistar-Kyoto and spontaneously hypertensive rats. The findings suggest that the expression of CYP4A3 and CYP4A8 may be critical to the early changes in eicosanoid formation and renal function in the young spontaneously hypertensive rat.

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Section: Cyp4a Expression and Activity In The Shr Kidney 369mentioning

confidence: 69%

Developmentally Regulated Expression of theCYP4AGenes in the Spontaneously Hypertensive Rat Kidney

et al. 1997

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“…2). Of equal interest is a 13-residue peptide (positions 307-319) that appears to be wholly conserved around position 315 in family 4 (12)(13)(14)(15)(26)(27)(28)(29)(30).…”

Section: Resultsmentioning

confidence: 99%

“…Comparison of the 511-amino acid Blaberus P450 with other P450s revealed highest similarity (32-36% positional identity) with family 4 proteins, a group of microsomal enzymes that primarily catabolize fatty acids and prostaglandins (10)(11)(12)(13)(14)(15)(16). The most striking region of homology was a 13-amino acid peptide found in all family 4 sequences examined but in no other P450.…”

mentioning

confidence: 95%

Cytochrome P450 family 4 in a cockroach: molecular cloning and regulation by regulation by hypertrehalosemic hormone.

Bradfield

Lee

Keeley

1991

Proc. Natl. Acad. Sci. U.S.A.

115

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Hypertrehalosemic hormone (a carbohydrate-mobilizing neuroendocrin decapepde) and starvation markedly increased levels of a cockroach (Blaberus discoidalis) fat body cytochrome P450 message. The gene represented by the cloned P450 cDNA has been named CYP4C1 (cytochrome P450 family 4, subfamily C, gene 1), a newly identified member of the ubiquitous cytochrome P450 monooxygenase gene superfamily. Blaberus CYP4C1 (511 amino acids, Mr = 58,485) has a hydrophobic NH2 terminus and a sequence near the COOH terminus that is homologous to the cysteine-containing heme-binding region definitive ofcytochromes P450. The cockroach sequence is 32-36% identical to mammalian family 4A and 4B enzymes. It contains a 13-residue sequence characteristic of family 4 but not other P450s. This study suggests that CYP4C1 is hormonally regulated in association with energy substrate mobilization and supports the idea that family 4 is an old and widespread gene family.The cytochrome P450 gene (CYPt) superfamily encodes NAD(P)H-dependent, heme-containing monooxygenases that metabolize numerous endogenous and exogenous substrates such as steroids, fatty acids, drugs, carcinogens, and pesticides (2,3). According to amino acid sequence analysis, =150 genes from the CYP superfamily have been assigned to 27 families (1), and a phylogeny of the superfamily has been proposed (1,4,5). Most P450 sequences have been obtained from vertebrates (especially mammals) but bacterial, yeast, fungal, invertebrate [including one insect (6)], and plant sequences are also described. The fact that cytochromes P450 are ubiquitous argues for descent of these proteins from a sequence that predated divergence of prokaryotes and eukaryotes.We have identified a cytochrome P450 sequence (deduced from cDNA) from the neotropical cockroach Blaberus discoidalis.t The cDNA was obtained from a fat body (analogous to vertebrate liver) library by differential hybridization designed to select gene sequences regulated by the hypertrehalosemic hormone (HTH) (7). HTH is a neuroendocrine decapeptide produced by the corpora cardiaca-paired neurosecretory glands attached to the brain. A major action for HTH is the stimulation of fat body glycogenolysis for the production of precursors for synthesis of trehalose, the main circulating carbohydrate of insects (8). HTH is a member of a large group of structurally related neuropeptides-the adipokinetic hormone/red pigment-concentrating hormone family (9)-that regulate energy substrate mobilization and metabolism in arthropods. Levels of the P450 mRNA are low in the fat body of decapitated (gland-free) adult male cockroaches, but strongly stimulated in decapitated animals by HTH administration. The P450 message is also stimulated by starvation in the presence but not in the absence of the head.Comparison of the 511-amino acid Blaberus P450 with other P450s revealed highest similarity (32-36% positional identity) with family 4 proteins, a group of microsomal enzymes that primarily catabolize fatty acids and prostaglandins (10-16). Th...

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“…She returned with the four clones of the cytochrome P450 4A gene family from rabbit lung and kidney, with which she expressed and purified the wild type and several chimeric constructs of these enzymes from Escherichia coli. Our collaboration with the Scripps group continued to be productive and critical to understanding the similarities and differences among these Ͼ85% sequence-identical cytochromes P450 (48,49). Dr. Ramani Narayanasami also joined us in San Antonio as a postdoctoral fellow and continued studies that he had begun at the Medical College of Wisconsin to determine the source of the reported "covalently bound" phosphate found in CYPOR by another group.…”

Section: Returning To Direct a Research Laboratorymentioning

confidence: 97%

“…While Scott Muerhoff was completing his Ph.D. requirements, I was learning molecular biological techniques during a mini-sabbatical with Dr. Eric Johnson at The Scripps Research Institute in La Jolla. This 3-month stint at Scripps initiated a collaboration to which members of our laboratory contributed enzymatic studies upon the cloning and expression of three cytochromes P450 from rabbit kidney, the first of the -hydroxylases to be cloned (48,49). During my sabbatical, I received an invitation from Dr. Merle Olson, the Chair of Biochemistry at The University of Texas Health Science Center at San Antonio, to examine the position of the first Robert A. Welch Foundation Endowed Chair at that institution.…”

Section: Chairing a Biochemistry Department?mentioning

confidence: 99%

A Professional and Personal Odyssey

Masters

2009

Journal of Biological Chemistry

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A s I began to write this article reflecting on my professional career in biomedical research, many memories of people, activities, situations, and experiences were swirling in my head. Because of the extraordinary support that I have received from my family, they are never far from my thoughts. I believe that sharing a few of these memories will be helpful for those who find themselves in similar situations or periods of professional development. This is not meant to be a comprehensive review of the fields to which I have had the privilege of contributing but a personal stroll down a path in which I have found extraordinary pleasure and from which I have derived a sense of accomplishment. The Journey BeginsI was born just prior to World War II into a middle class family in the historic town of Lexington, VA. My father was a radio announcer, a nightclub singer (he had a beautiful baritone voice), and the chosen master of ceremonies for most of the local shows in several cities in Southwest Virginia. Because my mother was a full-time parent, he had to hold several jobs to pay the bills. Because he had small children and was working in a vital industry (public information), he was not drafted during the war. Neither he nor my mother was college-educated, although both were well read and surprisingly cosmopolitan to have been raised in southwestern Virginia. Because I was his firstborn and my sister came along 2 years later, all of his dreams were wrapped up in what we would accomplish in our lifetimes. One of his oft-repeated admonitions was, "I don't care what you become as an adult, as long as you do your job well." However, it never escaped my attention that he wanted us to become professionals. Hence, the fact that we lived next door to a physician, his wife, and two boys had the inevitable effect that we observed that their somewhat higher grade of lifestyle (they owned their home, but we rented our two-bedroom apartment) was achievable. Both of my parents wanted desperately for their daughters to obtain college educations and they both insisted that we could "be anything we wanted to be." Unfortunately, I was not born with the artistic talent with which my sister and my brother (who was to arrive 22 years later during my first year in graduate school) were endowed, so my talents had to be sought elsewhere.In those days, we were known as "tomboys," and there was little hope for our interests to be more feminine, despite my mother's attempts, because our only playmates were boys on either side of us. We loved to climb trees, especially the cherry tree between our houses, and to play football and baseball and, best of all, cowboys and Indians. When my parents were vehemently against my getting a football for Christmas, I told my grandmother, and she made sure it was among her gifts for me, much to my mother's dismay. I loved to collect things, and I particularly loved outdoor activities. By the time I reached high school age, I had read Sir Arthur Conan Doyle's "The Adventures of Sherlock Holmes"; I admired Hol...

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Cloning and expression of three rabbit kidney cDNAs encoding lauric acid .omega.-hydroxylases

Cited by 62 publications

References 23 publications

Developmentally Regulated Expression of theCYP4AGenes in the Spontaneously Hypertensive Rat Kidney

Developmentally Regulated Expression of theCYP4AGenes in the Spontaneously Hypertensive Rat Kidney

Cytochrome P450 family 4 in a cockroach: molecular cloning and regulation by regulation by hypertrehalosemic hormone.

A Professional and Personal Odyssey

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