Immunological characterization of human liver α-D-mannosidase

Antiserum was raised against purified human liver α-D-mannosidase B. It precipitated α-mannosidases A and B from solution, demonstrating the close structural resemblance of these 2 forms of acidic α-mannosidase activity. A continuous enzymically active precipitin line with no spurs was obtained when α-mannosidase A and B were placed in adjacent wells on Ouchterlony double-diffusion plates. The antiserum precipitated acidic but not neutral α-mannosidase from an extract of human liver, confirming that the acidic and neutral activities are not closely related. Acidic activity was also precipitated from extracts of human brain, kidney and leucocytes by the antiserum. However, it did not cross-react with bovine acidic α-mannosidase activity or with the activity in human plasma that has an optimum pH of 5.5. The two acidic forms of human liver α-mannosidase, A and B, are immunologically identical but distinct from neutral α-mannosidase and that activity with an optimum pH of 5.5.

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The immunological characterization of several human ribonucleases by using primary binding tests

Biochemical Journal ◽

10.1042/bj1630419 ◽

1977 ◽

Vol 163 (3) ◽

pp. 419-426 ◽

Cited By ~ 18

Author(s):

E A Neuwelt ◽

M Schmukler ◽

M S Niziak ◽

P B Jewett ◽

C C Levy

Keyword(s):

Human Plasma ◽

Cross Reaction ◽

The Other ◽

Human Pancreas ◽

Human Tissues ◽

Antigenic Difference ◽

Immunological Characterization ◽

Similarities And Differences

RNAases (ribonucleases), purified from four human tissues, as well as bovine pancreatic RNAase (RNAase A), were studied by immunodiffusion methods and by two different primary binding tests. The enzymes fell into two groups immunologically, those purified from plasma and pancreas in one and those from spleen and liver in the other. No antigenic cross-reaction between the two groups was detected by any of the immunoassays used. There was a slight antigenic cross-reaction between the human and bovine pancreatic RNAases. The liver and spleen RNAases were immunologically identical by all criteria used, whereas a small but consistent antigenic difference between the human plasma and human pancreas enzymes was detected. The significance of this difference between the human plasma and pancreas RNAases is discussed in relation to similarities and differences in their properties.

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Characterization of antibody against human liver guanase by immunoblotting and immunohistochemical staining of human liver guanase by a direct labeling antibody-enzyme method.

Journal of Histochemistry & Cytochemistry ◽

10.1177/37.5.2649557 ◽

1989 ◽

Vol 37 (5) ◽

pp. 611-615 ◽

Cited By ~ 4

Author(s):

S Ito ◽

A Iwasaki ◽

J Syundo ◽

Y Tamura ◽

S Kishi ◽

...

Keyword(s):

Liver Enzyme ◽

Specific Antibody ◽

Immunohistochemical Staining ◽

Human Liver ◽

Liver Extract ◽

Precipitin Line ◽

Tissue Sections ◽

Immunohistochemical Reaction ◽

Enzyme Method

Human liver guanase was purified and a specific antibody against it was raised in rabbits. The antiserum formed a single precipitin line with human liver extract, and also completely inhibited the activity of the liver enzyme. An immunoblotting study showed that the antibody bound specifically to one band of protein with guanase activity and not to other proteins. Therefore, we concluded that this antiserum against the liver enzyme was suitable for use in immunohistochemical demonstration of guanase. In tissue sections, the immunohistochemical reaction with this antibody was positive in the same locations as the histochemical guanase reaction with DAB (3,3'-diaminobenzidine tetrahydrochloride).

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Biochemical and immunological characterization of three binding sites on human plasma fibronectin with different affinities for heparin

Biochemistry ◽

10.1021/bi00286a019 ◽

1983 ◽

Vol 22 (17) ◽

pp. 4113-4119 ◽

Cited By ~ 46

Author(s):

Leslie I. Gold ◽

Blas Frangione ◽

Edward Pearlstein

Keyword(s):

Human Plasma ◽

Binding Sites ◽

Plasma Fibronectin ◽

Immunological Characterization

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Solubilization and characterization of pellet-associated human brain α-L-fucosidase activity

Biochemical Journal ◽

10.1042/bj2290679 ◽

1985 ◽

Vol 229 (3) ◽

pp. 679-685 ◽

Cited By ~ 3

Author(s):

R L Hopfer ◽

J A Alhadeff

Keyword(s):

Human Brain ◽

Isoelectric Focusing ◽

Human Liver ◽

Gel Filtration ◽

Compelling Evidence ◽

Supernatant Fluid ◽

Ph Optimum ◽

Triton X ◽

Soluble Supernatant

The pellet-associated portion of human brain alpha-L-fucosidase (which represents approx. 20% of the homogenate activity) was solubilized with 0.5% (w/v) Triton X-100, characterized with regard to several properties and compared with the corresponding properties of the soluble supernatant-fluid enzyme in an attempt to find a second alpha-L-fucosidase in human brain. The solubilized and soluble alpha-L-fucosidase activities exhibited complete stability after storage at 2-4 degrees C for up to 29 days, comparable thermostability after preincubation at 50 degrees C, comparable apparent Km values (0.07-0.08 mM) for 4-methylumbelliferyl alpha-L-fucopyranoside, comparable hydrophobicity, comparable isoelectric-focusing profiles (six major forms, with pI values between 4.5 and 5.8) and comparable immunoprecipitation curves (with the IgG fraction of antisera prepared against human liver alpha-L-fucosidase). Differences in three properties were found between solubilized and soluble alpha-L-fucosidase activities: the solubilized activity was less stable to storage at −20 degrees C, had a 0.5-pH-unit neutral shift in its pH optimum (6.0) and had smaller Mr forms after gel filtration on Sephadex G-200. The overall results indicate that the pellet-associated and soluble portions of human brain alpha-L-fucosidase are quite similar in most of their properties. Thus there is still no compelling evidence for the existence of a second mammalian alpha-L-fucosidase.

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