A low-molecular-mass GTP-binding protein in the cytosol of germinated wheat embryos.
A low-molecular-mass protein able to bind GTP in both native and SDS-denaturating conditions was detected in the cytosol of embryos from wheat (Triticum aestivum L.) seeds germinated for 40 h. The protein fulfilled most of the distinguishing criteria common to eukaryotic small GTP-binding proteins. It retained the ability to bind GTP after SDS/PAGE and nitrocellulose blotting. The protein eluted from Sephadex G-200 gel filtration with a Ve/Vo value corresponding to a molecular mass of 18 kDa, whereas on SDS/PAGE the molecular mass was 20 kDa. The native protein, which showed an intrinsic GTPase activity highly sensitive to NaF, bound the guanine nucleotide with high specificity and with a relatively high affinity (Kd approximately 85 nM). The GTP-binding protein was not detectable in other subcellular fractions; in the microsomal fraction, two other peptides of low molecular mass (23.5 and 21.5 kDa) with GTP-binding activity were detected. These results indicate that in the cytosolic fraction of germinating wheat embryos there is a 20-kDa protein which is biochemically similar to the known small GTP-binding proteins that currently have been detected almost exclusively in the membrane fraction of plant material.
Dipartimento di Fisiologia delle Piante Coltivate e Chimica Agraria, Milano, Italy.
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Sacchi
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Pirovano
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Lucchini
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Cocucci
D002458:Cell Fractionation; D002850:Chromatography, Gel; D003593:Cytoplasm / Q000737:chemistry; D004591:Electrophoresis, Polyacrylamide Gel; D020558:GTP Phosphohydrolases / Q000378:metabolism; D019204:GTP-Binding Proteins / Q000737:chemistry* / Q000302:isolation & purification; D018525:Germination / Q000502:physiology; D006160:Guanosine Triphosphate / Q000378:metabolism; D008970:Molecular Weight; D011485:Protein Binding; D014908:Triticum / Q000737:chemistry* / Q000196:embryology
D006160:Guanosine Triphosphate; D020558:GTP Phosphohydrolases; D019204:GTP-Binding Proteins