Multiple forms of serum cholinesterase: Molecular weights of the isoenzymes

1970 ◽  
Vol 136 (2) ◽  
pp. 448-451 ◽  
Author(s):  
Rudolph V. LaMotta ◽  
Charles L. Woronick ◽  
Ralph F. Reinfrank
Keyword(s):  
Serum Cholinesterase ◽  
Multiple Forms ◽  
Molecular Weights

scholarly journals Multiple forms of acetylcholinesterase from human erythrocytes

1973 ◽  
Vol 133 (3) ◽  
pp. 521-527 ◽  
Author(s):  
David L. Wright ◽  
David T. Plummer
Keyword(s):  
Enzyme Activity ◽  
Electrophoretic Pattern ◽  
Human Erythrocytes ◽  
Serum Cholinesterase ◽  
Main Peak ◽  
Multiple Forms ◽  
Molecular Weights ◽  
Enzyme Substrate ◽  
Triton X ◽  
Naphthyl Acetate

1. Acetylcholinesterase from human erythrocytes was solubilized with Triton X-100 in strong salt solution and partially purified by (NH4)2SO4 fractionation. This preparation showed three main bands of enzyme activity after electrophoresis on polyacrylamide gel and incubation with either α-naphthyl acetate or acetylthiocholine as enzyme substrate. Two of the multiple forms were completely inhibited by 10μm-eserine and one only partially. Treatment with neuraminidase had no effect on the electrophoretic pattern; therefore sialic acid does not appear to determine or affect the ratios of the acetylcholinesterase multiple forms, unlike those of the serum cholinesterase. 2. Chromatography of the preparation on Sephadex G-200 revealed one major peak of enzyme activity and a suggestion of two minor zones of mol.wt. 546000, 184000 and 93000 (i.e. in the proportion 6:2:1). The main peak was almost completely separated from the Triton X-100 and the overall purification was about 600-fold. Further attempts to purify the enzyme by absorption on calcium phosphate gels were unsuccessful. 3. Electrophoresis of the enzyme preparation on a polyacrylamide gradient for 24h revealed three main bands that corresponded to the three values for molecular weights obtained by column chromatography. After 70h of electrophoresis a further three zones of activity developed making six molecular entities, the molecular weights of which were simple multiples of a monomer, thus resembling the cholinesterase found in serum.

scholarly journals Characterization of Multiple forms of Human Thrombin

1977 ◽  
Author(s):  
J.J. Gorman ◽  
P.A. Castaldi
Keyword(s):  
Specific Activity ◽  
Peptide Mapping ◽  
Multiple Forms ◽  
Bovine Thrombin ◽  
Molecular Weights ◽  
Human Thrombin ◽  
Affinity Resin ◽  
C 50 ◽  
High Degree

Human thrombin was obtained by activation of partially purified human prothrombin with venom of the Australian Taipan (oxyuranus scutellatus scutellatus).The crude thrombin was precipitated with ammonium sulphate and subsequently purified by chromatography on Sephadex G-75 CM-Sephadex C-50 and the affinity resin am inobenzamidine-CH-Sepharose. The final preparation had a specific activity of 1700 units per absorbance unit (A| cm 280n m Was herterogenous as shown by urea-acrylamide gel electrophoresis at acid pH and by isoelectric focusing. SDS-acrylamide electrophoresis revealed molecular weights of 39,000, 28,000, 25-23,000 and 15-12,000 for these proteins. The 39,000 dalton species predominated (greater than 90%) when the enzyme was inhibited with phenyImethanesuI phony I fluoride prior to dialysis against 0.02M sod i urn phosphate (pH 8.0) containing 0.1% SDS. Lack of such inhibition reduced the amount of the 39,000 dalton species to less than 60% with concomitant increase in the smaller species. Increase in the smaller species also occurred during incubation in 0.IM NaCI-0.I M Tris buffer (pH 8.0).Peptide mapping studies indicated that the smaller species were structurally related to the 39,000 dalton species. Amino acid compositions of tryptic peptides indicated a high degree of homology with bovine thrombin.It has been established that human thrombin can exist in at least two secondary structural forms of different molecular weights, probably due to autolytic degradation of the largest (39,000 dalton) protein species.

Production of multiple forms of glucoamylase in Aspergillus awamori

1987 ◽  
Vol 65 (8) ◽  
pp. 762-765 ◽  
Author(s):  
Resham S. Bhella ◽  
Illimar Altosaar
Keyword(s):  
Molecular Weight ◽  
Northern Blot Analysis ◽  
De Novo ◽  
In Vitro Translation ◽  
Aspergillus Awamori ◽  
Multiple Forms ◽  
Molecular Weights ◽  
Specific Mrnas

The biosynthesis of glucoamylases in Aspergillus awamori was studied by in vivo protein labelling and analysis of glucoamylase-specific mRNAs. Two types of glucoamylases with molecular weights of 100 000 and 82 000 were shown to be synthesized de novo. Deglycosylation of the 100 000 molecular weight glucoamylase type resulted in the formation of another glucoamylase form with molecular weight of about 94 000. De novo synthesis of two types of glucoamylases was further confirmed by the existence of two types of glucoamylase-specific mRNAs, as demonstrated by in vitro translation and Northern blot analysis studies.

scholarly journals Multiple forms of uridine kinase in normal and neoplastic rat liver

1971 ◽  
Vol 124 (5) ◽  
pp. 943-947 ◽  
Author(s):  
Gerald Krystal ◽  
Thomas E. Webb
Keyword(s):  
Rat Liver ◽  
Total Activity ◽  
Multiple Forms ◽  
Molecular Weights ◽  
Adult Rat ◽  
Uridine Kinase ◽  
Stable Species ◽  
Foetal Rat ◽  
Predominant Form ◽  
Slow Growing

Two species of uridine kinase with molecular weights of approximately 120000 (I) and 30000 (II) have been identified in the rat liver system. Species I predominates in the 7-day postnatal and adult rat liver and increases in the regenerating remnant of the latter after partial hepatectomy; the concentration of species II is low in these tissues. Species I also predominates in the slow-growing hepatomas 5123D and 7800. In contrast, II is the predominant form in the foetal rat liver and accounts for 40% of the total activity in the rapidly growing Novikoff ascites hepatoma. In contrast to species II, which was stable, species I was inactivated by preincubation for 30min at 37°C, before assay at 23°C.

Purification and Properties of Potato Phosphorylase Isozymes

1976 ◽  
Vol 31 (7-8) ◽  
pp. 424-432 ◽  
Author(s):  
Kirumakki N. Shivaram
Keyword(s):  
Starch Synthesis ◽  
Soluble Starch ◽  
Potato Tubers ◽  
Multiple Forms ◽  
Ph Optimum ◽  
Molecular Weights ◽  
Purification And Properties ◽  
Glucan Phosphorylase ◽  
Sulfhydryl Compounds ◽  
Synthesis And Degradation

Abstract Two multiple forms of α-glucan phosphorylase which migrate about half way in polyacryl-amide-gel electrophoresis (named “slow“ and “fast” isozyme), were isolated by combined chromatography and preparative electrophoresis after freezing the tissue from freshly harvested and from sprouting potato tubers respectively. Depending on the primer used for the synthesis reaction their pH optimum varied between 5.2 and 6.0 and the optimum temperature was 30 and 35 °C. The isoelectric point for the slow isozyme was at pH 5.0±0.1 and for the fast isozyme, pH 5.5±0.1, the molecular weights were 209 000±10 000 and 165 000±5000 and for their subunits 104 000±4000 and 40 000±2000 respectively. Both isozymes were inhibited by Hg2+, Ag+ and p-chloromercuro-benzoate (p-CMB). Fe2+ ions inhibited them partially. Mg2+, Mn2+ and sulfhydryl compounds activated both. Km values for the slow and fast isozymes with glucose-l-phosphate in presence of soluble starch was 6.7 and 8.0 mM, of amylose 14.3 and 20.0 mм and of glycogen 22.2 and 40.0 mм respectively. The affinity of the primer for the slow and the fast isozymes were as follows: soluble starch 0.5 and 1.0 mм, amylose 2.6 and 3.8 mм, glycogen 6.2 and 7.7 mм respectively. Km values of phosphorolysis with soluble starch was 0.8 and 0.5 mм, with amylose was 3.1 and 1.1 mм, and with glycogen was 6.5 and 1.3 mм respectively. As substrate and primer the soluble starch was superior and the glycogen was inferior. Amylose was in between. Kinetic parameters suggested the existence of α-glucan phosphorylase isozymes with different specificities: the slow one being more active in the direction of starch synthesis and the fast isozyme degrading faster the polyglucans. These observations suggest that the polyglucan synthesis and degradation in potato tubers may be regulated by the change in the proportion of slow and fast isozymes.

Isolation and purification of proteinase inhibitors from developing embryos of Hyalomma dromedarii

1990 ◽  
Vol 55 (2) ◽  
pp. 564-574 ◽  
Author(s):  
Ragaa R. Hamed ◽  
Mahmoud A. Ibrahim ◽  
Mamdouh Y. Kamel
Keyword(s):  
Embryonic Development ◽  
Proteinase Inhibitors ◽  
Deae Cellulose ◽  
Multiple Forms ◽  
Sharp Drop ◽  
Chymotrypsin Inhibitor ◽  
Isolation And Purification ◽  
Molecular Weights ◽  
High Level ◽  
Chymotrypsin Inhibitors

The changes in the level of trypsin, chymotrypsin, subtilisin and papain inhibitors were examined during embryonic development of H. dromedarii. The high level of trypsin and chymotrypsin inhibitors in the oviposited eggs did not change during embryogenesis and was followed by a sharp drop after hatching. On the other hand, the papain inhibitor exhibited very low level throughout embryonic development and was followed by a significant increase in the newly hatched larvae. A purification scheme for trypsin and chymotrypsin inhibitors which involved chromatography on DEAE-cellulose, cellulose phosphate and Ultrogel AcA 54 was established. Using synthetic substrates six multiple forms of proteinase inhibitors with different molecular weights and properties were isolated. They were classified into three specific trypsin inhibitors F1a, F4a, and F4b, one specific chymotrypsin inhibitor F1b and to mixed trypsin-chymotrypsin inhibitors F2 and F3. Although treatment that may cause fragmentation of the native inhibitors were avoided during purification, multiple forms of trypsin and chymotrypsin inhibitor were isolated. The significancy of such multiplicity is not understood.

scholarly journals Purification and properties of urease from bovine rumen

1977 ◽  
Vol 163 (3) ◽  
pp. 495-501 ◽  
Author(s):  
S Mahadevan ◽  
F D Sauer ◽  
J D Erfle
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Maximum Velocity ◽  
Maximum Activity ◽  
Rumen Bacteria ◽  
Multiple Forms ◽  
Bovine Rumen ◽  
Molecular Weights ◽  
Purification And Properties ◽  
High Concentrations

Urease (urea amidohydrolase, EC 3.5.1.5) was extracted from the mixed rumen bacterial fraction of bovine rumen contents and purified 60-fold by (NH4)2SO4 precipitation, calcium phosphate-gel adsorption and chromatography on hydroxyapatite. The purified enzyme had maximum activity at pH 8.0. The molecular weight was estimated to be 120000-130000. The Km for urea was 8.3 X 10(-4) M+/-1.7 X 10(-4) M. The maximum velocity was 3.2+/-0.25 mmol of urea hydrolysed/h per mg of protein. The enzyme was stabilized by 50 mM-dithiothreitol. The enzyme was not inhibited by high concentrations of EDTA or phosphate but was inhibited by Mn2+, Mg2+, Ba2+, Hg2+, Cu2+, Zn2+, Cd2+, Ni2+ and Co2+. p-Chloromercuribenzenesulfphonate and N-ethylmaleimide inhibited the enzyme almost completely at 0.1 mM. Hydroxyurea and acetohydroxamate reversibly inhibited the enzyme. Polyacrylamide-gel electrophoresis showed that the mixed rumen bacteria produce ureases which have identical molecular weights and electrophoretic mobility. No multiple forms of urease were detected.

Multiple forms of serum cholinesterase

1968 ◽  
Vol 124 ◽  
pp. 299-305 ◽  
Author(s):  
R.V. LaMotta ◽  
R.B. McComb ◽  
C.R. Noll ◽  
H.J. Wetstone ◽  
R.F. Reinfrank
Keyword(s):  
Serum Cholinesterase ◽  
Multiple Forms

scholarly journals Comparative Studies on Multiple Forms of Serum Cholinesterase in Various Species

1987 ◽  
Vol 36 (2) ◽  
pp. 199-204 ◽  
Author(s):  
Satoshi UNAKAMI ◽  
Sachiko SUZUKI ◽  
Etsuko NAKANISHI ◽  
Kazuaki ICHINOHE ◽  
Mariko HIRATA ◽  
...  
Keyword(s):  
Comparative Studies ◽  
Serum Cholinesterase ◽  
Multiple Forms

Multiple forms of somatostatin-like immunoreactivity in the hypothalamus and amygdala of the rat: selective localization of somatostatin-28 in the median eminence

1985 ◽  
Vol 105 (3) ◽  
pp. 383-389 ◽  
Author(s):  
A. R. Pierotti ◽  
A. J. Harmar
Keyword(s):  
Median Eminence ◽  
High Performance ◽  
Hormone Secretion ◽  
Growth Hormone Secretion ◽  
Gel Filtration ◽  
Multiple Forms ◽  
Gel Filtration Chromatography ◽  
Molecular Weights ◽  
Hypothalamic Regulation ◽  
Composite Peak

ABSTRACT The presence of multiple forms of somatostatin-like immunoreactivity (SSLI) in the rat hypothalamus was confirmed using a sensitive radioimmunoassay in conjunction with gel filtration chromatography and high performance liquid chromatography (HPLC). Gel filtration chromatography of hypothalamic extracts revealed the presence of four forms of SSLI with estimated molecular weights of 1500, 3000, 6000 and 10000. Analysis by HPLC indicated that the 1500 and 3000 mol. wt forms of SSLI corresponded respectively to somatostatin-14 (SS14) and somatostatin-28 (SS28) whereas the 6000 and 10 000 mol. wt forms eluted together as a composite peak of high molecular weight somatostatin (HMW-SS). The proportions of SS14 (63%), SS28 (12%) and HMW-SS (25%) present in the hypothalamus were similar to those in the amygdala (59, 9 and 32% respectively). In contrast, the median eminence contained a greater proportion of SS28 than the other tissues: SS14, SS28 and HMW-SS were present in the proportions 40:24:26%. These results show that the rat median eminence differs from the hypothalamus as a whole in containing SS14 and SS28 in almost equimolar concentrations. The localized abundance of SS28 in the nerve terminals of the median eminence suggests a specific role for this peptide in the hypothalamic regulation of growth hormone secretion. J. Endocr. (1985) 105, 383–389

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