scholarly journals Purification and partial characterization of human polyamine synthases

1989 ◽  
Vol 259 (3) ◽  
pp. 879-886 ◽  
Author(s):  
E O Kajander ◽  
L I Kauppinen ◽  
R L Pajula ◽  
K Karkola ◽  
T O Eloranta
Keyword(s):  
Gel Electrophoresis ◽  
Cross Reactivity ◽  
Specific Pattern ◽  
Native Enzyme ◽  
Spermidine Synthase ◽  
Enzyme Preparations ◽  
Spermine Synthase ◽  
Apparent Homogeneity ◽  
Gradient Gel Electrophoresis ◽  
Pore Gradient

Spermidine synthase was purified to apparent homogeneity from human spleens (8700-fold) by affinity chromatography. The native enzyme was composed of two subunits of identical Mr (35,000) and showed an apparent Mr of 62,000 in pore-gradient gel electrophoresis. Its pI was 5.1, Spermine synthase was purified to apparent homogeneity from placenta (5300-fold) and from kidney (4600-fold). The native enzyme was composed of two subunits of identical Mr (45,000) and showed an apparent Mr of 78,000 in pore-gradient gel electrophoresis. In isoelectric focusing it revealed two bands, with pI values of 4.9 and 5.0. Both synthases were present in all human tissues studied, but revealed a clear tissue-specific pattern. Mouse antisera against spermidine synthase revealed only one band, of Mr 35,000, in all purified enzyme preparations and in crude human tissue extracts in immunoblotting. Antisera against spermine synthase showed an immunoreactive band corresponding to the Mr of the subunit of spermine synthase. These antisera did not indicate any cross-reactivity in immunoblotting. Thus spermine synthase and spermidine synthase do not share homologous antigenic sites and are totally different proteins.

scholarly journals Affinity-chromatographic purification of S-adenosyl-l-homocysteine hydrolase. Some properties of the enzyme from rat liver

1981 ◽  
Vol 193 (2) ◽  
pp. 503-512 ◽  
Author(s):  
E O Kajander ◽  
A M Raina
Keyword(s):  
Rat Liver ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Adenosine Kinase ◽  
Native Enzyme ◽  
Apparent Homogeneity ◽  
A Subunit ◽  
Adenosine Derivatives ◽  
Pore Gradient

S-Adenosyl-L-homocysteine hydrolase has been purified to apparent homogeneity from rat liver by means of affinity chromatography on 8-(3-aminopropylamino)adenosine linked to Sepharose. The purified enzyme was free from adenosine kinase and adenosine deaminase activities and was homogeneous on SDS/polyacrylamide-gel electrophoresis which gave a subunit mol.wt. of 47 000. The native enzyme showed some microheterogeneity on polyacrylamide-gel electrophoresis under increased-resolution conditions but was homogeneous on isoelectric focusing (pI 5.6). The molecular weight of the native enzyme was about 220 000 as judged by pore-gradient electrophoresis. The native enzyme bound adenosine tightly and showed Km values of 0.6 microM, 0.9 microM and 60 microM for adenosine, S-adenosyl-L-homocysteine and L-homocysteine respectively. The enzyme was rapidly inactivated when incubated in the presence of adenosine, S-adenosyl-L-homocysteine or several adenosine derivatives or analogues. Inactivation took place both at 0 and 37 degrees C. Freezing in the absence of glycerol resulted in the appearance of dissociation products of the oligomeric protein. Multimer formation was observed at low thiol concentrations.

Antibody to spermine: a natural biological constituent

Science ◽  
1980 ◽  
Vol 208 (4448) ◽  
pp. 1178-1181 ◽  
Author(s):  
D Bartos ◽  
F Bartos ◽  
RA Campbell ◽  
DP Grettie ◽  
P Smejtek
Keyword(s):  
Affinity Chromatography ◽  
Gel Electrophoresis ◽  
Binding Constants ◽  
Cross Reactivity ◽  
Polyamine Metabolism ◽  
Regulatory Function ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Immunodiffusion Test ◽  
Gradient Gel Electrophoresis

A protein that binds spermine specifically was separated from normal rabbit serum by affinity chromatography. Immunoelectrophoresis, the Ouchterlony immunodiffusion test, and gradient gel electrophoresis indicated that this protein has immunoglobulin characteristics and consists of several populations of antibodies to spermine. These were sequentially released from Sepharose-spermine gel by step-wise elution with solutions ranging in pH from 4 to 1. The binding constants varied from 5.0 x 10(8) to 11.1 x 10(8) liters per mole. These globulins did not react with monoacetylputrescine, L-ornithine, L-lysine, and histamine. Negligible cross-reactivity was detected with spermidine, putrescine, N8-monoacetylspermidine, cadaverine, and diaminopropane. Since perturbations in polyamine metabolism have been identified in several diseases, the study of extracellular polyamine homeostasis may reveal an important regulatory function for this protein.

Transverse pore gradient gel electrophoresis, using the PhastSystem

1994 ◽  
Vol 15 (1) ◽  
pp. 1028-1031 ◽  
Author(s):  
Zsuzsanna Buzás ◽  
David L. Wheeler ◽  
Mark M. Garner ◽  
Dietmar Tietz ◽  
Andreas Chrambach
Keyword(s):  
Gel Electrophoresis ◽  
Gradient Gel Electrophoresis ◽  
Pore Gradient
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