麻彤辉
博士研究生 教授 博士生导师
吉林大学 第二医院
膜蛋白结构与功能;高通量筛选药物发现;
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- 姓名:麻彤辉
- 目前身份:在职研究人员
- 担任导师情况:博士生导师
- 学位:
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学术头衔:
国家“百千万”人才工程国家级人选, 国家杰出青年科学基金获得者, 享受国务院特殊津贴专家
- 职称:高级-教授
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学科领域:
细胞生物学
- 研究兴趣:膜蛋白结构与功能;高通量筛选药物发现;
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946
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成果数
14
【期刊论文】Defective Secretion of Saliva in Transgenic Mice Lacking Aquaporin-5 Water Channels*
麻彤辉, Tonghui Ma, Yualin Song, Annemarie Gillespie, Elaine J. Carlson, Charles J. Epstein, and A. S. Verkman‡
THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 274, No.29, Issue of July 16, pp. 20071-20074, 1999,-0001,():
-1年11月30日
Aquaporin-5 (AQP5) is a water-selective transporting protein expressed in epithelial cells of serous acini in salivary gland. We generated AQP5 null mice by targeted gene disruption. The genotype distribution from intercross of founder AQP5 heterozygous mice was 70: 69: 29 wild type: heterozygote: knockout, indicating impaired prenatal survival of the null mice. The knockout mice had grossly normal appearance, but grew; 20% slower than litter-matched wild-type mice when placed on solid food after weaning. Pilocarpine-stimulated saliva production was reduced by more than 60% in AQP5 knockout mice. Compared with the saliva from wildtype mice, the saliva from knockout mice was hypertonic (420 mosM) and dramatically more viscous. Amylase and protein secretion, functions of salivary mucous cells, were not affected by AQP5 deletion. Water channels AQP1 and AQP4 have also been localized to salivary gland; however, pilocarpine stimulation studies showed no defect in the volume or composition of saliva in AQP1 and AQP4 knockout mice. These results implicate a key role for AQP5 in saliva fluid secretion and provide direct evidence that high epithelial cell membrane water permeability is required for active, near-isosmolar fluid transport.
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麻彤辉, Tonghui Ma, Baoxue Yang, Michael A. Matthay, and A. S. Verkman
Am. J. Respir. Cell Mol. Biol. Vol. 19, pp. 143-149, 1998,-0001,():
-1年11月30日
T1 a is a protein of unknown function that is expressed at the plasma membrane in epithelia involved in fluid transport, including type I alveolar epithelial cells, choroid plexus, and ciliary epithelium. The purpose of this study was to test the hypothesis that T1 a functions as a water channel or a regulator of aquaporin-type water channels that are coexpressed with T1 a. Two complementary DNAs (cDNAs)(hT1a-1 and hT1 a-2) encoding human isoforms of T1 a were cloned by homology to the rat T1 a coding sequence. The cDNAs encoded 164 (hT1 a-1) and 162 (hT1 a -2) amino acid proteins with high homology to rat T1 a in a putative membrane-spanning domain. hT1 a-1 transcripts of 2.6 and 1.4 kb were detected in human lung, heart, and skeletal muscle, and a single hT1 a-2 transcript of 1.2 kb was detected in human lung. Rat and mouse T1 a were isolated by reverse transcription-polymerase chain reaction and confirmed by DNA sequence analysis. Expression of mouse, rat, and human T1 a isoforms in Xenopus oocytes did not increase osmotic water permeability (Pf) above that in water-injected oocytes, nor was there an effect of protein kinase A or C activation; Pf was increased. 10-fold in positive control oocytes expressing aquaporin (AQP)1 or AQP5. Coexpression of AQP1 or AQP5 with excess T1a gave P f not different from that in oocytes expressing AQP1 or AQP5 alone. Oocyte plasma membrane localization of epitope-tagged T1 a protein was confirmed and quantified by immunoprecipitation of microdissected plasma membranes. Quantitative densitometry indicated that the single-channel water permeability of T1 a is under 23102 16cm 3/s, suggesting that T1 a is not involved in the high transalveolar water permeability in intact lung. The cloning of hT1 a isoforms may permit the development of an assay of type I cell antigen in airspace fluid as a marker of human lung injury. Ma, T., B. Yang, M. A. Matthay, and A. S. Verkman. 1998. Evidence gainst a role of mouse, rat, and two cloned human T1 a isoforms as a water channel or a regulator of aquaporin-type water channels. Am. J. Respir. Cell Mol. Biol. 19: 143-149.
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麻彤辉, Tonghui Ma‡, L. Vetrivel‡, Hong Yang‡, Nicoletta Pedemonte§, Olga Zegarra-Moran§, Luis J. V. Galietta§, and A. S. Verkman‡¶
THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 277, No.40, Issue of October 4, pp. 37235-37241, 2002,-0001,():
-1年11月30日
and were CFTR-selective, reversible, and nontoxic. Several compounds, the most potent being a trifluoromethylphenylbenzamine, activated the CF-causing mutant G551D, but with much weaker affinity (Kd>10M). When added for 10min, none of the compounds activated Phe508-CFTR in transfected cells grown at 37℃ (with Phe508-CFTR trapped in the endoplasmic reticulum). However, after correction of trafficking by 48h of growth at 27℃, tetrahydrocarbazol and N-phenyltriazine derivatives strongly stimulated Cl conductance with Kd<1M. The new activators identified here may be useful in defining molecular mechanisms of CFTR activation and as lead compounds in CF drug development.
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麻彤辉, Tonghui Ma, Baoxue Yang, Annemarie Gillespie, Elaine J. Carlson, Charles J. Epstein, and A.S. Verkman
Volume 100, Number 5, September 1997, 957-962,-0001,():
-1年11月30日
Aquaporin-4 (AQP4) is a mercurial-insensitive, water-selective channel that is expressed in astroglia and basolateral plasma membranes of epithelia in the kidney collecting duct, airways, stomach, and colon. A targeting vector for homologous recombination was constructed using a 7-kb SacI AQP4 genomic fragment in which part of the exon 1 coding sequence was deleted. Analysis of 164 live births from AQP4[+/-] matings showed 41 [-/-], 83 [+/-], and 40 [-/-] genotypes. The [-/-] mice expressed small amounts of a truncated AQP4 transcript and lacked detectable AQP4 protein by immunoblot analysis and immunocytochemistry. Water permeability in an AQP4-enriched brain vesicle fraction in [+/+] mice was high and mercurial insensitive, and was decreased by 14-fold in [-/-] mice. AQP4 deletion did not affect growth or tissue morphology at the light microscopic level. Northern blot analysis showed that tissue-specific expression of AQPs 1, 2, 3, and 5 was not affected by AQP4 deletion. Maximum urine osmolality after a 36-h water deprivation was (in mosM, n=15) [+/+] 3,34
water transport, aquaporin, AQP4, mercurial-insensitive water channel, urinary concentration),
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【期刊论文】Impaired Stratum Corneum Hydration in Mice Lacking Epidermal Water Channel Aquaporin-3*
麻彤辉, Tonghui Ma‡§, Mariko Hara‡¶, Rachid Sougrat, Jean-Marc Verbavatz, and A. S. Verkman‡
THE JOURNAL OF BIOLOGICAL CHEMISTRY Vol. 277, No.19, Issue of May 10, pp. 17147-17153, 2002,-0001,():
-1年11月30日
The water and solute transporting properties of the epidermis have been proposed to be important determinants of skin moisture content and barrier properties. The water/small solute-transporting protein aquaporin-3 (AQP3) was found by immunofluorescence and immunogold electron microscopy to be expressed at the plasma membrane of epidermal keratinocytes in mouseskin. We studied the role of AQP3 in stratum corneum (SC) hydration by comparative measurements in wildtype and AQP3 null mice generated in a hairless SKH1 genetic background. The hairless AQP3 null mice had normal perinatal survival, growth, and serum chemistries but were polyuric because of defective urinary concentrating ability. AQP3 deletion resulted in a >4-fold reduced osmotic water permeability and >2-fold reduced glycerol permeability in epidermis. Epidermal, dermal, and SC thickness and morphology were not grossly affected by AQP3 deletion. Surface conductance measurements showed remarkably reduced SC water content in AQP3 null mice in the hairless genetic background (165
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麻彤辉, Tonghui Ma, Antonio Frigeri, Shih-Ting Tsai, Jean-Marc Verbavatzt, and A. S. Verkman
THE JOURNAL OF BIOLOGICACLH EMISTRY, Vol. 268, No.30, Issue of October 25, pp. 22756-22764, 1993,-0001,():
-1年11月30日
CHIP28 is a major water transporting protein in erythrocytes and plasma membranes in kidney proxi-mal tubule and thin descending limb of Henle. Chinese hamster ovary cells were stably transfected with the coding sequence of cloned rat kidney CHIP28k using expression vectors containing cytomegalovirus or Rous sarcoma virus promoters. Clonal cell populations expressed a 1.3-kilobase mRNA on Northern blot probed by CHIP28k cDNA and a 28-kDa protein on immunoblot probed by a polyclonal CHIP28 antibody. The clone with greatest expression produced -8 X 10’copies of CHIP28k protein/cell. Plasma membrane os-motic water permeability (P f), measured by stopped-flow light scattering, was 0.004cm/s in control (vec-tor-transfected) cells (10C) and 0.014cm/s in the CHIP28k-transfected cells. P f in CHIP28k-transfected cells had an activation energy of 4.9 kcal/mol and was reversibly inhibited by HgC12. CHIP28k expression did not affect the transport of protons and the small polar non-lectrolytes urea and formamide. CHIP28k im-munoreactivity and function was then determined in subcellular fractions. P f in 6-carboxyfluorescein-la-beled endocytic vesicles, measured by a stopped-flow fluorescence quenching assay, was 0.002cm/s (control cells) and 0.011cm/s (CHIP28k-transfected cells); Pt in transfected cells was inhibited by HgC12. Immuno-blotting of fractionated endoplasmic reticulum, Golgi, and plasma membranes revealed high densities of CHIP28k (-5000 monomers/pm2 in plasma membrane) with different glycosylation patterns; functional water transport activity was present only in Golgi and plasma membrane vesicles. Antibody detection of CHIP28k by confocal fluorescence microscopy and immunogold electron microscopy revealed localization to plasma membrane and intracellular vesicles. These studies es-tablish a stably transfected somatic cell line that strongly expresses functional CHIP28k water chan-nels. As in the original proximal tubule cells, the ex-pressed CHIP28k protein is a selective water channel that is functional in endocytic vesicles and the cell plasma membrane.
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【期刊论文】Lung fluid transport in aquaporin-5 knockout mice
麻彤辉, Tonghui Ma, Norimasa Fukuda, Yuanlin Song, Michael A. Matthay, and A.S. Verkman
The Journal of Clinical Investigation|January 2000|Volume 105|Number 1,-0001,():
-1年11月30日
The mammalian lung expresses water channel aquaporin-1 (AQP1) in microvascular endothelia, AQP4 in airway epithelia, and AQP5 at the apical plasma membrane in type I cells of alveolar epithelia. We previously studied the role of AQP1 and AQP4 in lung fluid transport using knockout mice. Here, we examined the role of AQP5 using AQP5 knockout mice, which were recently shown to manifest defective saliva secretion. AQP5 deletion did not affect lung morphology at the light microscopic level, nor did it affect the distribution or expression of aquaporins 1, 3, or 4. Airspace-capillary osmotic water permeability (Pf) was measured in isolated perfused lungs by pleural surface fluorescence and gravimetric methods. Pf was reduced 10-fold by AQP5 deletion and was further reduced by 2- to 3-fold in AQP1/AQP5 double-knockout mice. Hydrostatic lung edema in response to acute increases in pulmonary artery pressure was not affected by AQP5 deletion. Active alveolar fluid absorption was measured in an in situ lung model from the increase in concentration of a volume marker in an isosmolar alveolar instillate. Interestingly, fluid absorption did not differ in litter-matched AQP5 knockout mice, nor was there an effect of AQP5 deletion when fluid absorption was maximally stimulated by pretreatment of mice with keratinocyte growth factor. These results indicate that AQP5 is responsible for the majority of water transport across the apical membrane of type I alveolar epithelial cells. The unimpaired alveolar fluid clearance in AQP5-null mice indicates that high alveolar water permeability is not required for active, near-isosmolar fluid transport.
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麻彤辉, Tonghui Ma, , Jay R. Thiagarajah, Hong Yang, Nitin D. Sonawane, Chiara Folli, Luis J.V. Galietta, and A.S. Verkman
The Journal of Clinical Investigation|December 2002|Volume 110|Number 11,-0001,():
-1年11月30日
Secretory diarrhea is the leading cause of infant death in developing countries and a major cause of morbidity in adults. The cystic fibrosis transmembrane conductance regulator (CFTR) protein is required for fluid secretion in the intestine and airways and, when defective, causes the lethal genetic disease cystic fibrosis. We screened 50,000 chemically diverse compounds for inhibition of cAMP/flavone-stimulated Cl-transport in epithelial cells expressing CFTR. Six CFTR inhibitors of the 2-thioxo-4-thiazolidinone chemical class were identified. The most potent compound discovered by screening of structural analogs, CFTRinh-172, reversibly inhibited CFTR short-circuit current in less than 2 minutes in a voltage-independent manner with KI approximately 300nM. CFTRinh-172 was nontoxic at high concentrations in cell culture and mouse models. At concentrations fully inhibiting CFTR, CFTRinh-172 did not prevent elevation of cellular cAMP or inhibit non-CFTR Cl-channels, multidrug resistance protein-1 (MDR-1), ATP-sensitive K+ channels, or a series of other transporters. A single intraperitoneal injection of CFTRinh-172 (250
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【期刊论文】Nephrogenic diabetes insipidus in mice lacking aquaporin-3 water channels
麻彤辉, Tonghui Ma, Yuanlin Song, Baoxue Yang, Annemarie Gillespie, Elaine J. Carlson, Charles J. Epstein, and A. S. Verkman*
4386-4391/PNAS/April 11, 2000/vol. 97/no.8,-0001,():
-1年11月30日
Aquaporin-3 (AQP3) is a water channel expressed at the basolateral plasma membrane of kidney collecting-duct epithelial cells. The mouse AQP3 cDNA was isolated and encodes a 292-amino acid wateryglycerol-transporting glycoprotein expressed in kidney, large airways, eye, urinary bladder, skin, and gastrointestinal tract. The mouse AQP3 gene was analyzed, and AQP3 null mice were generated by targeted gene disruption. The growth and phenotype of AQP3 null mice were grossly normal except for polyuria. AQP3 deletion had little effect on AQP1 or AQP4 protein expression but decreased AQP2 protein expression particularly in renal cortex. Fluid consumption in AQP3 null mice was more than 10-fold greater than that in wild-type litter mates, and urine osmolality (<275 milliosmol) was much lower than in wild-type mice (>1,200 milliosmol). After 1-desamino-8-D-arginine-vasopressin administration or water deprivation, the AQP3 null mice were able to concentrate their urine partially to 30% of that in wild-type mice. Osmotic water permeability of cortical collecting-duct basolateral membrane, measured by a spatial filtering optics method, was >3-fold reduced by AQP3 deletion. To test the hypothesis that the residual concentrating ability of AQP3 null mice was due to the inner medullary collecting-duct water channel AQP4, AQP3yAQP4 double-knockout mice were generated. The double-knockout mice had greater impairment of urinary-concentrating ability than did the AQP3 single-knockout mice. Our findings establish a form of nephrogenic diabetes insipidus produced by impaired water permeability in collecting-duct basolateral membrane. Basolateral membrane aquaporins may thus provide blood-accessible targets for drug discovery of aquaretic inhibitors.
water transport AQP3 u kidney urinary-concentrating mechanism polyuria
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麻彤辉, Tonghui Ma, Antonio FrigeriS, Hajime HasegawaS, and A. S. Verkman
THE JOURNAL OF BIOLOGIGAL CHENISTRY, Vol. 286, No.84, Jssue of August 26, pp. 21845-21849, 1994.,-0001,():
-1年11月30日
In searching for a basolateral membrane water trans-porter in rat kidney with homology to channel formingintegral protein (CHIP28), water channel-collecting duct (WCH-CD), and mercurial-insensitive water chan-nel (MIWC), we cloned a new member of the major in-trinsic protein family (GLIP, -cero1 Intrinsic B o -tein). GLIP cDNA had an 865-base pair open reading frame encoding a 30.6-kDa protein with 19-23% amino acid identity to the water channels and 36% identity to the bacterial glycerol facilitator GlpF. Northern blot analysis showed a 6.6-kilobase mFWA encoding GLIP in kidney, brain, and lung; RT-PCWSouthern blot analysis indicated expression of GLIP in kidney, brain, lung, eye, colon, stomach, and skeletal muscle, but not in heart, liver, and spleen. In situ hybridization in rat kidney showed GLIP mRNA expression in medullary collecting duct. Immunofluorescence with a peptide-derived poly-clonal antibody showed GLIP protein expression in ba-solateral membrane of kidney collecting duct principal cells and brain meningeal cells. Functional measure-ments in Xenopus oocytes expressing GLIP cRNA showed a>2O-fold increase in [SHlglycerol uptake com-pared with water-iqjected oocytes; glycerol uptake was inhibited 88% by diisothiocyanodisdfonic stilbene (0.2mm) and 36% by phloretin (0.25mm). GLIP did not func-tion as a transporter for water, urea, inositol, glucose, lactate, and monovalent ions. Glycerol uptake in oocytes expressing CHIP28 and MIWC was not different from that in water-injected controls. GLIP represents the first mammalian water channel homolog that selectively transports a solute other than water. The physiological substrate(s) and role(s) of GLIP remain to be elucidated.
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