Purification of the low-molecular-weight form of adenosine deaminase from rabbit intestine

Galactocerebrosidase (β-d-galactosyl-N-acylsphingosine galactohydrolase; EC 3.2.1.46) activity of brain and liver preparations from normal individuals and patients with Krabbe disease (globoid-cell leukodystrophy) have been separated by gel filtration into four different molecular-weight forms. The apparent mol.wts. were 760000±34000 and 121000±10000 for the high- and low-molecular-weight forms (peaks I and IV respectively) and 499000±22000 (mean±s.d.) and 256000±12000 for the intermediate forms (peaks II and III respectively). On examination by sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, the high- and low-molecular-weight forms revealed a single protein band with a similar mobility corresponding to a mol.wt. of about 125000. Antigenic identity was demonstrated between the various molecular-weight forms of the normal and the mutant galactocerebrosidases by using antisera against either the high- or the low-molecular-weight enzymes. The high-molecular-weight form of galactocerebrosidase was found to possess higher specific activity toward natural substrates when compared with the low-molecular-weight form. It is suggested that the high-molecular-weight enzyme is the active form in vivo and an aggregation process that proceeds from a monomer (mol.wt. approx. 125000) to a dimer (mol.wt. approx. 250000) and from the dimer to either a tetramer (mol.wt. approx. 500000) or a hexamer (mol.wt. approx. 750000) takes place in normal as well as in Krabbe-disease tissues.

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Synthesis of a Low-Molecular-Weight Form of Exopolysaccharide by Bradyrhizobium japonicum USDA 110

Applied and Environmental Microbiology ◽

10.1128/aem.67.2.1011-1014.2001 ◽

2001 ◽

Vol 67 (2) ◽

pp. 1011-1014 ◽

Cited By ~ 10

Author(s):

Heather A. Louch ◽

Karen J. Miller

Keyword(s):

Molecular Weight ◽

Bradyrhizobium Japonicum ◽

Compositional Analysis ◽

Mass Spectrometry Analysis ◽

Low Molecular Weight ◽

Nuclear Magnetic Resonance Analysis ◽

Spectrometry Analysis ◽

Resonance Analysis ◽

Dimeric Form ◽

Molecular Weight Form

ABSTRACT A novel extracellular low-molecular-weight polysaccharide was detected as a contaminant within extracellular cyclic β-1,6-β-1,3-glucan preparations from Bradyrhizobium japonicum USDA 110 cultures. Compositional analysis, methylation analysis, and nuclear magnetic resonance analysis revealed that this low-molecular-weight polysaccharide was composed of the same pentasaccharide repeating unit previously described for the high-molecular-weight form of the exopolysaccharide (EPS) synthesized by B. japonicum strains. Mass spectrometry analysis indicated that the size of this low-molecular-weight form of EPS was consistent with a dimeric form of the pentasaccharide repeating unit.

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Immunoaffinity Chromatography of Canine High-Molecular-Weight Renin: Partial Purification and Characterization

Clinical Science ◽

10.1042/cs0650117 ◽

1983 ◽

Vol 65 (2) ◽

pp. 117-120 ◽

Cited By ~ 2

Author(s):

Fumihiko Ikemoto ◽

Victor J. Dzau ◽

Edgar Haber ◽

Kazuo Takaori ◽

Kenjiro Yamamoto

Keyword(s):

Molecular Weight ◽

High Molecular Weight ◽

Specific Activity ◽

Immunoaffinity Chromatography ◽

Partial Purification ◽

Low Molecular Weight ◽

Specific Method ◽

Disulphide Bonds ◽

Binding Substance ◽

Molecular Weight Form

1. Canine high-molecular-weight renin (mol. wt. 60 000) is believed to be a complex of renin (low-molecular-weight form, mol. wt. 40 000) and renin-binding substance. The immunocross-reactivity of high-molecular-weight renin and low-molecular-weight renin was demonstrated by using antibodies specific to low-molecular-weight renin. 2. Immunoaffinity chromatography with renin-specific antibodies coupled to Sepharose provided a simple and specific method for isolation of high-molecular-weight renin. High-molecular-weight renin with a specific activity of 137 600 ng of ANG I h−1 mg−1 of protein (19.6 Goldblatt units/mg of protein) was obtained. 3. This high-molecular-weight renin was stable in dithiothreitol (25 mmol/l), suggesting that disulphide bonds may not be involved in the binding mechanism between low-molecular-weight renin and renin-binding substance. 4. However, exposure to low pH (3.0) resulted in conversion of high-molecular-weight renin into the low-molecular-weight form.

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Preliminary crystallographic study on a low molecular weight form of bacterial plasminogen activator staphylokinase

Acta Crystallographica Section D Biological Crystallography ◽

10.1107/s090744499700084x ◽

1997 ◽

Vol 53 (4) ◽

pp. 480-481 ◽

Cited By ~ 1

Author(s):

D. Chattopadhyay ◽

J. E. Stewart ◽

C. D. Smith ◽

L. J. DeLucas ◽

S. V. L. Narayana

Keyword(s):

Molecular Weight ◽

Plasminogen Activator ◽

Low Molecular Weight ◽

Crystallographic Study ◽

Molecular Weight Form

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Studies on the appearance of a hepatic copper-binding protein in normal and zinc-deficient rats

British Journal Of Nutrition ◽

10.1079/bjn19760061 ◽

1976 ◽

Vol 36 (1) ◽

pp. 101-112 ◽

Cited By ~ 50

Author(s):

I. Bremner ◽

N. T. Davies

Keyword(s):

Molecular Weight ◽

Metal Binding ◽

Binding Protein ◽

Gel Filtration ◽

Apparent Molecular Weight ◽

Low Molecular Weight ◽

Copper Binding ◽

Hepatic Copper ◽

Copper Binding Protein ◽

Molecular Weight Form

1. A study has been made by gel-filtration techniques of the soluble copper- and zinc-binding proteins in rat liver after both intraperitoneal injection of Cu and dietary Cu supplementation.2. Liver Cu and Zn concentrations increased after injection of Cu, both metals accumulating in the cytosol, mainly in a fraction with an apparent molecular weight of (about 12 000)3. When Zn-deficient rats were injected with Cu, there was little change in liver Zn concentration and the occurrence of Cu in the low-molecular-weight form (about 12 000) was more transient. At most periods after injection, Cu accumulated mainly in a fraction with a molecular weight greater than 65 000.4. When the rats were Cu-loaded by dietary supplementation, virtually no Cu or Zn was found in the low-molecular-weight form in Zn-deficient rats, although they were found in the Zn-supplemented animals.5. The results suggest that Zn is essential for the accumulation of Cu in this form, but not for Cu to stimulate production of the metal-binding protein by a process requiring active protein synthesis.

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