Preparation and characterisation of monoclonal and polyclonal antibodies to maize membrane auxin-binding protein

Napier, Richard; Venis, Michael A.; Bolton, Michael; Richardson, Louise; Butcher, G. W.

doi:10.1007/bf00397659

Cited by 94 publications

(83 citation statements)

References 20 publications

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“…The polyclonal antibody against ABP1, but not the preimmune antiserum, immunoprecipitates a 22-kD protein photolabeled with [3H]5-N3IAA. The recognition of the 22-kD band by these antibodies comigrated with recognition by antibodies previously described by Napier et al (23). Furthermore, antisera used by Napier et al (23) The level of IAA and IAA esters in the apical mesocotyl was determined by GC-SIM-MS using '3C6-IAA as an internal standard as described by Chen et al (7), and it was found that 2 h after continuous irradiation the free IAA level was approximately 50% of the dark control tissue (Fig.…”

Section: Materials and Methods Plant Tissue Chemicals And Light Treamentioning

confidence: 72%

See 1 more Smart Citation

Red Light-Regulated Growth

Jones¹,

Cochran²,

Lamerson³

et al. 1991

Plant Physiol.

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We examined the changes in the levels of indoleacetic acid (IAA), IAA esters, and a 22-kilodalton subunit auxin-binding protein (ABP1) in apical mesocotyl tissue of maize (Zea mays L.) during continuous red light (R) irradiation. These changes were compared with the kinetics of R-induced growth inhibition in the same tissue. Upon the onset of continuous irradiation, growth decreased in a continuous manner following a brief lag period. The decrease in growth continued for 5 hours, then remained constant at 25% of the dark rate. The abundance of ABP1 and the level of free IAA both decreased in the mesocotyl. Only the kinetics of the decrease in IAA within the apical mesocotyl correlated with the initial change in growth, although growth continued to decrease even after IAA content reached its final level, 50% of the dark control. This decrease in IAA within the mesocotyl probably occurs primarily by a change in its transport within the shoot since auxin applied as a pulse moved basipetally in Rirradiated tissue at the same rate but with half the area as dark control tissue. In situ localization of auxin in etiolated maize shoots revealed that R-irradiated shoots contained less auxin in the epidermis than the dark controls. Irradiated mesocotyl grew 50% less than the dark controls even when incubated in an optimal level of auxin. However, irradiated and dark tissue contained essentially the same amount of radioactivity after incubation in [14C]IAA indicating that the light treatment does not affect the uptake into the tissue through the cut end, although it is possible that a small subset of cells within the mesocotyl is affected. These observations support the hypothesis that R causes a decrease in the level of auxin in epidermal cells of the mesocotyl, consequently constraining the growth of the entire mesocotyl.R-causes the elongation rate of the cells in the apical 1 cm of the etiolated maize mesocotyl (e.g. 16, 29) and cell

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Section: Materials and Methods Plant Tissue Chemicals And Light Treamentioning

confidence: 72%

“…We have reexamined this problem by looking at the kinetics of the R-induced changes in IAA level, ABP1 level (18,23), and growth kinetics. Because there is some indirect evidence that ABP1 may mediate auxin effects on cell walls (e.g.…”

mentioning

confidence: 99%

Red Light-Regulated Growth

Jones¹,

Cochran²,

Lamerson³

et al. 1991

Plant Physiol.

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“…Control experiments with antibodies against a luminal epitope, i.e. the KDEL endoplasmic reticulum retention signal (30,31), showed that digitonin did not permeabilize the endoplasmic reticulum membrane. For the major hydrophilic loop domain, essentially identical results were obtained.…”

Section: Discussionmentioning

confidence: 98%

“…To analyze this question, we used low concentrations of digitonin to selectively permeabilize the plasma membrane (see "Materials and Methods") and antibodies against the amino-terminal domain (antiserum B14), against the Myc tag introduced at the amino terminus (mAb 9E10), against the second loop domain (antibody 519), and, finally, against the hydrophilic loop domain (antiserum B16) of PS1. To monitor the permeabilization procedure, rat monoclonal antibody against the luminal endoplasmic reticulum retention signal KDEL was used (30,31). Antibodies against the amino-terminal domain of PS1, against the amino-terminally inserted Myc tag (Fig.…”

Section: Cytoplasmic Orientation Of the Amino-terminal Domain And Hydmentioning

confidence: 99%

Phosphorylation, Subcellular Localization, and Membrane Orientation of the Alzheimer's Disease-associated Presenilins

Strooper¹,

Beullens²,

Contreras

et al. 1997

Journal of Biological Chemistry

272

187

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Presenilins 1 and 2 are unglycosylated proteins with apparent molecular mass of 45 and 50 kDa, respectively, in transfected COS-1 and Chinese hamster ovary cells. They colocalize with proteins from the endoplasmic reticulum and the Golgi apparatus in transfected and untransfected cells. In COS-1 cells low amounts of intact endogeneous presenilin 1 migrating at 45 kDa are detected together with relative larger amounts of presenilin 1 fragments migrating between 18 and 30 kDa. The presenilins have a strong tendency to form aggregates (mass of 100 -250 kDa) in SDS-polyacrylamide gel electrophoresis, which can be partially resolved when denatured by SDS at 37°C instead of 95°C. Sulfation, glycosaminoglycan modification, or acylation of the presenilins was not observed, but both proteins are posttranslationally phosphorylated on serine residues. The mutations Ala-246 3 Glu or Cys-410 3 Tyr that cause Alzheimer's disease do not interfere with the biosynthesis or phosphorylation of presenilin 1. Finally, using low concentrations of digitonin to selectively permeabilize the cell membrane but not the endoplasmic reticulum membrane, it is demonstrated that the two major hydrophilic domains of presenilin 1 are oriented to the cytoplasm. The current investigation documents the posttranslational modifications and subcellular localization of the presenilins and indicates that postulated interactions with amyloid precursor protein metabolism should occur in the early compartments of the biosynthetic pathway.

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“…LAMP and CD63 antibodies were obtained from the Developmental Studies Hybridoma Bank, University of Iowa (LAMP-1 clone H4A3, LAMP-2 clone H4B4, CD63 clone H5C6), anti-LBPA was a gift from J Gruenberg (Department of Biochemistry, University of Geneva, Switzerland), 35 anti-KDEL MAC256 was a gift from G Butcher (The Babraham Institute, Babraham, UK) 36 and anti-EEA1 and anti-Golgi GM130 were from BD Biosciences (Pharmingen). Secondary antibodies were from Jackson Immunoresearch Labs.…”

Section: Methodsmentioning

confidence: 99%