Biomedical subjects

Feng Yan

Publications and source records attributed to Feng Yan.

At least 55 records · Page 3Linked to original sources

PubMed2004

SV40 T antigen interacts with Nbs1 to disrupt DNA replication control.

Nijmegen breakage syndrome (NBS) is characterized by radiation hypersensitivity, chromosomal instability, and predisposition to cancer. Nbs1, the NBS protein, forms a tight complex with Mre11 and Rad50, and these interactions contribute to proper double-strand break repair. The simian virus 40 (SV40) oncoprotein, large T antigen (T), also interacts with Nbs1, and T-containing cells experience chromosomal hyperreplication in a manner dependent on T/Nbs1 complex formation. A substantial fraction of NBS-deficient fibroblasts reinitiate DNA replication in discrete regions, and wild-type Nbs1 corrects this defect. Similarly, synthesis of an N-terminal Nbs1 fragment induced DNA rereplication and tetraploidy, in NBS-deficient but not NBS-proficient cells. Moreover, SV40 origin-containing DNA hyperreplicated in T-containing NBS-deficient cells by comparison with T-containing, Nbs1-reconstituted derivatives. Thus, Nbs1 suppresses rereplication of cellular DNA and SV40 origin-containing replicons, and T targets Nbs1, thereby enhancing the yield of new SV40 genomes during viral DNA replication.

Acid Anhydride Hydrolases↗

PubMed2004

Cyclin-dependent kinases phosphorylate human Cdt1 and induce its degradation.

Eukaryotic cells tightly control DNA replication so that replication origins fire only once during S phase within the same cell cycle. Cell cycle-regulated degradation of the replication licensing factor Cdt1 plays important roles in preventing more than one round of DNA replication per cell cycle. We have previously shown that the SCF(Skp2)-mediated ubiquitination pathway plays an important role in Cdt1 degradation. In this study, we demonstrate that human Cdt1 is a substrate of Cdk2 and Cdk4 both in vivo and in vitro. Overexpression of cyclin-dependent kinase inhibitors such as p21 and p27 dramatically suppresses the phosphorylation of Cdt1, disrupts the interaction of Cdt1 with the F-box protein Skp2, and blocks the degradation of Cdt1. Further analysis reveals that Cdt1 interacts with cyclin/cyclin-dependent kinase (Cdk) complexes through a cyclin/Cdk binding consensus site, located at the N terminus of Cdt1. A Cdt1 mutant carrying four amino acid substitutions at the Cdk binding site dramatically reduces associations with cyclin/Cdk complexes. This mutant is not phosphorylated, fails to bind Skp2 and is more stable than wild-type Cdt1. These data suggest that cyclin/Cdk-mediated Cdt1 phosphorylation is required for the association of Cdt1 with the SCF(Skp2) ubiquitin ligase and thus is important for the cell cycle dependent degradation of Cdt1 in mammalian cells.

Binding Sites↗

PubMed2004

Correlation between serum vascular endothelial growth factor and endostatin levels in patients with breast cancer.

Serum vascular endothelial growth factor (VEGF) and endostatin levels were detected in 59 patients with breast cancer before surgery and at 3 weeks after surgery. Pre-operatively, their levels were significantly elevated and correlated with each other. Post-operatively, VEGF level decreased significantly and endostatin remained at a high level. Patients with both normalized VEGF and elevated endostatin following surgery had a lower risk of relapse than patients whose VEGF failed to normalize. Univariate and multivariate analyses showed a correlation between elevated VEGF level and short free-relapse survival. These findings suggest a new angiogenesis balance is formed in the patients after surgery and such a resultant balance may be beneficial for the prognosis of breast cancer, which deserves more extensive study.

Adult↗

PubMed2004

Noncompetitive enzyme immunoassay for carcinoembryonic antigen by flow injection chemiluminescence.

BACKGROUND: Recently, many automated immunoassay analyzers have been developed for carcinoembryonic antigen (CEA) to overcome the shortcomings in traditional immunoassay methods that are time-consuming and labor-intensive. Flow injection immunoassay (FIIA) has been increasingly applied to laboratory medicine due to its ease in automation, rapid speed and reproducible results. It is important to develop a FIIA method for CEA determination. METHODS: Based on a noncompetitive immunoassay format, a CEA-immobilized immunoaffinity column inserted in the flow system was used to trap the unbound horseradish peroxidase (HRP)-labeled antibody after an off-line incubation of CEA and HRP-labeled anti-CEA. The trapped enzyme conjugate was detected by injecting substrates to produce an enhanced chemiluminescence (CL). RESULTS: The linear range for CEA was 1.0-25 ng/ml with a correlation coefficient of 0.997 and a detection limit of 0.5 ng/ml. The sampling and chemiluminescence detection time for one sample was 5 min after a preincubation procedure of 25 min. Twenty five human serum samples detected by this method were in good agreement with the results obtained by immunoradiometric assay (IRMA). CONCLUSIONS: This method could be used for rapid analysis of CEA and potentially other antigens.

Carcinoembryonic Antigen↗

PubMed2004

Novel amperometric immunosensor for rapid separation-free immunoassay of carcinoembryonic antigen.

A novel immunosensor for rapid separation-free determination of carcinoembryonic antigen (CEA) in human serum is proposed. The immunosensor is prepared by co-immobilizing thionine and horseradish peroxidase (HRP)-labeled CEA antibody on a glassy carbon electrode (GCE) through covalently binding them to GCE with a glutaraldehyde (GA) linkage. The electrochemical behavior of the immobilized thionine displays a surface-controlled electrode process with an average electron transfer rate constant of 4.74+/-2.99 s(-1). It can be used as an electron transfer mediator for enzymatic activity detection of the HRP-labeled antibody to CEA. After the immunosensor is incubated with CEA solution at 23 degrees C for 40 min, the access of activity center of the HRP to thionine is partly inhibited, which leads to a linear decrease in the catalytic efficiency of the HRP to the oxidation of immobilized thionine by H(2)O(2) at -300 mV over two CEA concentration ranges from 0.5 to 3.0 and 3.0 to 167 ng/ml. Under optimal conditions, the detection limit for the CEA immunoassay is 0.1 ng/ml at three times background noise. The immunosensor shows good accuracy and acceptable storage stability, precision and reproducibility with intra-assay CVs of 6.1% and 5.8% at 2.5 and 50 ng/ml CEA, respectively, and an inter-assay CV of 6.3% at 50 ng/ml. This method is economical and shortens the analytical time, making it potentially attractive for clinical immunoassays.

Antibodies↗

PubMed2004

Chemiluminescent immunosensor for CA19-9 based on antigen immobilization on a cross-linked chitosan membrane.

A novel chemiluminescent immunosensor for carbohydrate antigen 19-9 (CA19-9) based on the immobilization of CA19-9 on the cross-linked chitosan membrane was developed. The different membranes were characterized by atomic force microscopy (AFM) and infrared spectrum, respectively. Based on a noncompetitive immunoassay format, this proposed chemiluminescent immunosensor enabled a low-cost, flexible and rapid determination for CA19-9 in combination with flow injection analysis (FIA). After an off-line incubation of the analyte CA19-9 with horseradish peroxidase (HRP)-labeled anti-CA19-9, the mixture was injected into the immunosensor, which led to the trapping of free HRP-labeled anti-CA19-9 by the immobilized antigen in the immunosensor. The trapped HRP-labeled antibody was detected by chemiluminescence due to its catalytic activity following the reaction of luminol and H2O2. Under optimal conditions, the decreased chemiluminescent signal of the immunosensor was proportional to the CA19-9 concentration in the range of 2.0-25 U/ml with a detection limit of 1.0 U/ml. The immunosensor showed an acceptable accuracy and good reproducibility. The results of 20 human serum samples detected by this method were in acceptable agreement with those obtained by immunoradiometric assay. The proposed immunosensor provided a new promising tool for practical clinical detection of the serum CA19-9 level.

Antibodies↗

PubMed2004

Noncompetitive enzyme immunoassay for alpha-fetoprotein using flow injection chemiluminescence.

A novel, direct noncompetitive flow injection enzyme immunoassay for alpha-fetoprotein (AFP) was developed by enhanced chemiluminescence detection. The method was based on off-line incubation of AFP and horseradish peroxidase (HRP)-labeled anti-AFP, and then trapping of the unbound enzyme conjugate by an immunoaffinity column filled with AFP-modified Sepharose. The immunocomplex formed in incubation passed through the column and then was directly detected by a postcolumn chemiluminescence technique. The optimal conditions for the immunoassay procedure and chemiluminescence detection were established. At a 1:10 dilution of enzyme conjugate solution, the linear range for chemiluminescence detection of AFP was from 2.0 to 75 ng/mL with a correlation coefficient of 0.993 and a coefficient of variation of 2.67% at 30 ng/mL. The detection limit was 0.5 ng/mL. This method was flexible, sensitive, and rapid. The immunoaffinity column of 200 microL could be repeatedly used 100 times without a single decrease. The whole assay time including the preincubation step was only 30 min for one sample.

Biomarkers↗

PubMed2004

[New developments in application of laser capture microdissection].

Isolation of pure targeted cells is an important and essential step for the molecular analysis of tissue lesion occurred in the progression of disease. Laser capture microdissection (LCM) is a novel technique that can be applied to obtain pure targeted cell subgroup or even a single cell quickly and precisely under the microscope, thus the problem of tissue heterogeneity in molecular analysis can be tackled successfully. In this article, the principles, advantages and disadvantages of LCM were introduced. New developments in the application of LCM were summarized in two fields (DNA analysis and gene expression analysis) respectively. Meanwhile, possible directions of the future developments of LCM were put forward.

Animals↗

PubMed2004

[Detection of WT1 expression in bone marrow of acute leukemia patients with real-time quantitative RT-PCR].

OBJECTIVE: To investigate Wilms' tumor gene (WT1) expression levels in bone marrow (BM) of acute leukemia patients (ALs). METHODS: A real-time quantitative reverse transcriptase polymerase chain reaction (RQ-RT-PCR) method was established for detecting WT1 and internal reference GAPDH expression levels in BM of 108 ALs and 23 non-leukemia controls by Light Cycler. RESULTS: The median expression levels of WT1 in 70 newly diagnosed ALs and 11 relapsed ALs were statistically higher than those in 23 ALs in complete remission (CR) and 23 non-leukemic controls (75.10 and 89.56 vs 2.07 and 1.51 respectively). No statistic differences was found between the CR group and control group, nor between the newly diagnosed group and relapsed group. Of the 70 newly diagnosed ALs, median WT1 expression level of acute granulocytic leukemias was significantly higher than that of acute monocytic leukemias (M(5)), but there was no statistic differences among the M(1), M(2), M(3) and ALL subtypes. Furthermore the WT1 levels were not correlated to peripheral WBC counts, BM blast percentage and multidrug resistant gene (mdr1) expression at presentation, but correlated to chromosome karyotypes. Dynamic analysis of WT1 levels of 2 patients on treatment showed that WT1 expression levels predicted relapse. CONCLUSION: WT1 expression levels in ALs were strikingly higher than that in non-leukemias. WT1 can be a marker for detecting MRD and evaluating therapy efficacy in leukemias.

Acute Disease↗

PubMed2003

Reagentless amperometric immunosensors based on direct electrochemistry of horseradish peroxidase for determination of carcinoma antigen-125.

A novel strategy for immunoassay and the preparation of reagentless immunosensors was proposed. This strategy was based on the immobilization of antigen and the direct electrochemistry of horseradish peroxidase (HRP) that was labeled to an antibody. A reagentless immunosensor for carcinoma antigen-125 (CA 125) determination was developed. The immunosensor was prepared by immobilizing CA 125 with titania sol-gel on a glassy carbon electrode by the vapor deposition method. The incubation of the immunosensor in phosphate buffer solution (PBS) including HRP-labeled CA 125 antibody led to the formation of a HRP-modified surface. The immobilized HRP displayed its direct electrochemistry with a rate constant of 3.04 +/- 1.21 s(-1). With a competition mechanism, a differential pulse voltammetric determination method for CA 125 was established by the peak current decrease of the immobilized HRP. The current decrease resulted from the competitive binding of the CA 125 in sample solution and the immobilized CA 125 to the limited amount of HRP-labeled CA 125 antibody. Under optimal conditions, the current decrease was proportional to CA 125 concentration ranging from 2 to 14 units mL(-1) with a detection limit of 1.29 units mL(-1) at a current decrease by 10%. The CA 125 immunosensor showed good accuracy and acceptable precision and fabrication reproducibility with intraassay CVs of 8.7 and 5.5% at 8 and 14 units mL(-1) CA 125 concentrations, respectively, and interassay CV of 19.8% at 8 units mL(-1). The storage stability was acceptable in a pH 7.0 PBS at 4 degrees C for 15 days. The proposed method provided a new promising platform for clinical immunoassay.

Antibodies, Monoclonal↗

PubMed2003

Biochemical and spectroscopic studies on (S)-2-hydroxypropylphosphonic acid epoxidase: a novel mononuclear non-heme iron enzyme.

The last step of the biosynthesis of fosfomycin, a clinically useful antibiotic, is the conversion of (S)-2-hydroxypropylphosphonic acid (HPP) to fosfomycin. Since the ring oxygen in fosfomycin has been shown in earlier feeding experiments to be derived from the hydroxyl group of HPP, this oxirane formation reaction is effectively a dehydrogenation process. To study this unique C-O bond formation step, we have overexpressed and purified the desired HPP epoxidase. Results reported herein provided initial biochemical evidence revealing that HPP epoxidase is an iron-dependent enzyme and that both NAD(P)H and a flavin or flavoprotein reductase are required for its activity. The 2 K EPR spectrum of oxidized iron-reconstituted fosfomycin epoxidase reveals resonances typical of S = (5)/(2) Fe(III) centers in at least two environments. Addition of HPP causes a redistribution with the appearance of at least two additional species, showing that the iron environment is perturbed. Exposure of this sample to NO elicits no changes, showing that the iron is nearly all in the Fe(III) state. However, addition of NO to the Fe(II) reconstituted enzyme that has not been exposed to O(2) yields an intense EPR spectrum typical of an S = (3)/(2) Fe(II)-NO complex. This complex is also heterogeneous, but addition of substrate converts it to a single, homogeneous S = (3)/(2) species with a new EPR spectrum, suggesting that substrate binds to or near the iron, thereby organizing the center. The fact that NO binds to the ferrous center suggests O(2) can also bind at this site as part of the catalytic cycle. Using purified epoxidase and (18)O isotopic labeled HPP, the retention of the hydroxyl oxygen of HPP in fosfomycin was demonstrated. While ether ring formation as a result of dehydrogenation of a secondary alcohol has precedence in the literature, these catalyses require alpha-ketoglutarate for activity. In contrast, HPP epoxidase is alpha-ketoglutarate independent. Thus, the cyclization of HPP to fosfomycin clearly represents an intriguing conversion beyond the scope entailed by common biological epoxidation and C-O bond formation.

Amino Acid Sequence↗

PubMed2003

Specific ubiquitin-conjugating enzymes promote degradation of specific nuclear receptor coactivators.

Nuclear receptor coactivators (NRCoAs) are nuclear hormone receptor-associated regulatory proteins that interact with members of the nuclear receptor superfamily in the presence of their cognate ligand, enhancing their transcriptional activity. The identification of ubiquitin-proteasome pathway proteins as coactivators provides evidence that ubiquitin-proteasome-mediated protein degradation plays an integral role in eukaryotic gene transcription. It has also been observed that nuclear receptors themselves are ubiquitinated and degraded in a hormone-dependent manner and that ubiquitin-proteasome function is essential for most nuclear receptors to function as transactivators. Here, we show that specific ubiquitin-proteasome pathway enzymes target specific NRCoA proteins in vivo and in vitro. First, using a temperature-sensitive cell line that contains a thermolabile ubiquitin-activating E1 enzyme, we confirmed that NRCoA proteins are targets of the ubiquitin-proteasome pathway. Then using coimmunoprecipitation studies, we also demonstrate that in vivo, NRCoA proteins are ubiquitinated. Finally, we illustrate that in vitro, NRCoA ubiquitination and degradation depend on the ubiquitin-activating enzyme (E1) and on specific ubiquitin-conjugating enzymes (E2) for each of the coactivators.

Animals↗

PubMed2003

Immunological assay for carbohydrate antigen 19-9 using an electrochemical immunosensor and antigen immobilization in titania sol-gel matrix.

We describe a novel electrochemical immunosensor for carbohydrate antigen 19-9 (CA19-9) based on the immobilization of CA19-9 with titania sol-gel on a graphite electrode (GE) by vapor deposition. The CA19-9 membrane was characterized using scanning electron microscopy and proved to be chemically clean, porous and homogeneous. The incubation of the immunosensor in a solution containing horseradish peroxidase (HRP)-labeled CA19-9 antibody led to the binding of HRP-labeled antibody with the immobilized antigen. The immobilized HRP catalyzed the oxidation of catechol by H(2)O(2) and this provided a competitive method for the measurement of serum CA19-9. The response current decreased with increasing CA19-9 concentration in the incubation solution. The effects of pH, amount of HRP-labeled antibody, incubation time and temperature were explored to provide optimum analytical performance. Under optimal conditions, the current decrease of the immunosensor was proportional to CA19-9 concentrations in the range of 3-20 U/ml with a detection limit of 2.68 U/ml at a current decrease of 10%. The detection of CA19-9 in two serum samples obtained from clinically diagnosed patients with pancreatic carcinoma showed acceptable accuracy. The proposed immunosensor provides a new promising tool for the clinical immunoassay of CA19-9.

Biosensing Techniques↗

PubMed2003

Using divisional history to measure hematopoietic stem cell self-renewal and differentiation.

OBJECTIVES: The purpose of this study was to investigate cell fates and long-term repopulating potential of a primitive hematopoietic stem cell (HSC) population (i.e., FR25Lin(-) cells) in vitro. MATERIALS AND METHODS: FR25Lin(-) cells were isolated by elutriation and cell sorting and cultured with a combination of cytokines for 7 days. Utilizing the membrane dye PKH-26, cultured cells were separated into two subsets based on their proliferation rates and assayed for progenitors and HSC. RESULTS: Fresh FR25Lin(-) cells were mostly quiescent; however, some of this population entered cell cycle after cytokine exposure reaching a peak 4 to 5 days after culture. Two subsets of cultured cells were isolated: 1) cells that had divided several times (PKH(dull) cells) and 2) cells that remained undivided or divided only once or twice (PKH(bright) cells). The PKH(dull) cells accounted for 94% of total viable cells in culture after 5 days. The PKH(dull) subset contained all the multi-potential in vivo progenitors (CFU-S) and 10 times more committed progenitors (CFU-C). Quantitative analysis of HSC engraftment from the PKH(bright) subset demonstrated stem cell maintenance. For the PKH(dull) subset, on day 5, HSC numbers increased. By day 7, increased differentiation in the PKH(dull) population supports expanding differentiation divisions. CONCLUSIONS: Our primitive HSC population underwent different types of cell divisions stimulated by cytokines, resulting in subsets with different self-renewal and differentiation potentials. This in vitro/in vivo model provides a useful tool for studies of early events during HSC self-renewal and differentiation.

Animals↗

PubMed2003

Metabolic effects of a novel bioartificial liver on serum from severe hepatitis patients: an in vitro study.

OBJECTIVE: To establish a novel bioartificial liver (BAL) consisting of spheroids of porcine hepatocytes in a hollow-fiber bioreactor, and to perform an in vitro study on its metabolic effects on the serum from severe hepatitis B patients. METHODS: Hepatocytes were isolated from pup pigs and cultured as aggregate spheroids through rotation and vibration. Phase-contrast microscopy, transmission electron microscopy, and scanning electron microscopy were used for morphological detection of hepatocyte spheroids. The hepatocyte spheroids were then transferred into the shell of a polysulfone hollow-fiber bioreactor, creating a novel BAL. Diluted serum samples of severe hepatitis B patients were circulated for 3 hours each into the bioreactor, by using an extracorporeal circulatory system. Every half hour, including both before and after perfusion, serum samples were collected to assay total bilirubin (TBIL), total protein (TP), albumin (ALB), and globulin (GLB) concentrations in order to judge the metabolic effects of this novel BAL. RESULTS: Most hepatocytes had formed spheroids with high viability after 24 hours in culture. After 3 hours of perfusion, when compared with the control group, the serum concentration of TBIL in the treatment group decreased significantly (P < 0.01), but the serum concentrations of TP and ALB increased significantly (P < 0.05). CONCLUSIONS: Hepatocytes can be conveniently cultured as aggregate spheroids through a rotation and vibration method. The novel BAL is efficient in removing bilirubin from the serum of severe hepatitis B patients, and in supplying the serum with ALB. Thus, the BAL might provide effective therapy for patients with severe hepatitis B.

Bilirubin↗

PubMed2003

[Expression of TGF-beta receptor II in periretinal membranes of proliferative vitreoretinopathy].

PURPOSE: To investigate the expression of transforming growth factor-beta receptor II (TGF-beta R II) in periretinal membranes of proliferative vitreoretinopathy (PVR). METHODS: Immunohistochemistry and in situ hybridization were used to detect the expression of TGF-beta R II protein and mRNA in sixteen proliferative membranes of thirteen PVR patients. RESULTS: Immunohistochemically staining showed that membranes labeled negative "-", weak positive "+", positive "2+" and strong positive staining "3+" were respectively noted one, one, three and four membranes in nine C2-C3 membranes. Total positive cell rate is 88.9%. Among seven D1-D3 membranes, one membrane was labeled "-", three were "+" and another three membranes were labeled "2+". Total positive cell rate is 85.7%. Positive staining cells were of a type of epithelial-like cells which frequently found pigment in or around them. Relation between TGF-beta R II expression and membrane grades appeared no correlation (P > 0.05). Results of in situ hybridization were almost consistent with that of immunohistochemistry. CONCLUSIONS: Both TGF-beta R II protein and mRNA are highly expressed and up-regulated in PVR membranes by cytokine stimulation, moreover TGF-beta may play important role in the pathogenesis of PVR.

Adolescent↗

PubMed2002

[Site-directed mutagenesis of atrial natriuretic peptide gene and effect of the mutations on its diuretic activity in nephrotic rats].

OBJECTIVE: To prolong the half-life and enhance the biological activity of the human atrial natriuretic peptide (hANP), a peptide hormone, which is synthesized and released mainly by cardiac atrial myocytes and possesses potent natriuretic, diretic, and vasorelaxant properties. METHODS: The site-directed mutation technique based on polymerase chain reaction was performed to get the mutant of the human ANP gene (mhANP), and the retroviral expression vector, pLHY24, in which mhANP gene is under the transcriptional control of the human cytomegalovirus promoter, was constructed. The naked plasmid DNA of pLHY24 and positive control vector, pLHY19, in which the wild-type hNAP gene is in the same conditions as mhANP gene in pLHY24, and negative control vector, pLNCX without purpose gene, at a dose of 5 mg/kg body weight was injected intramuscularly into the rats with experimental renal disorder induced with adriamycin (ADR), respectively. RESULTS: DNA sequencing result proved that the respected mhANP gene with the point mutations of TTC(131)/Phe-->TCC/Ser and ATG(135)/Met-->ATA/Ile has been obtained. In comparison with negative control group (87 +/- 7.1 pg/ml), a single intramuscular injection of expression vector harboring mhANP or hANP gene resulted in an obvious increase in plasma level of mhANP (107 +/- 7.8 pg/ml, t = 4.65, P < 0.01) or hANP (113 +/- 8.6 pg/ml, t = 5.71, P < 0.01) 5 days after injection. A significant elevation in the ratio of urine volume to body weight was occurred after both of mhANP gene and hANP gene delivery as compared with negative control and the effect lasted for more than 15 days. The diuretic activity of mhANP gene delivery was 1.6-, 2.0-, and 1.9-fold higher than that of hANP gene 5, 10, and 15 days after gene transfer, respectively. However, there were no statistical differences in the concentrations of K(+) and Na(+) in urine. CONCLUSIONS: Both of mhANP and hANP gene delivery into the rats with experimental nephropathy could improve their directic function obviously and the diuretic activity of the former is stronger than that of the latter significantly.

Animals↗

PubMed2002

Adaptation of H+-pumping and plasma membrane H+ ATPase activity in proteoid roots of white lupin under phosphate deficiency.

White lupin (Lupinus albus) is able to adapt to phosphorus deficiency by producing proteoid roots that release a huge amount of organic acids, resulting in mobilization of sparingly soluble soil phosphate in rhizosphere. The mechanisms responsible for the release of organic acids by proteoid root cells, especially the trans-membrane transport processes, have not been elucidated. Because of high cytosolic pH, the release of undissociated organic acids is not probable. In the present study, we focused on H+ export by plasma membrane H+ ATPase in active proteoid roots. In vivo, rhizosphere acidification of active proteoid roots was vanadate sensitive. Plasma membranes were isolated from proteoid roots and lateral roots from P-deficient and -sufficient plants. In vitro, in comparison with two types of lateral roots and proteoid roots of P-sufficient plants, the following increase of the various parameters was induced in active proteoid roots of P-deficient plants: (a) hydrolytic ATPase activity, (b) Vmax and Km, (c) H+ ATPase enzyme concentration of plasma membrane, (d) H+-pumping activity, (e) pH gradient across the membrane of plasmalemma vesicles, and (f) passive H+ permeability of plasma membrane. In addition, lower vanadate sensitivity and more acidic pH optimum were determined for plasma membrane ATPase of active proteoid roots. Our data support the hypothesis that in active proteoid root cells, H+ and organic anions are exported separately, and that modification of plasma membrane H+ ATPase is essential for enhanced rhizosphere acidification by active proteoid roots.

Adaptation, Physiological↗