Hexokinase A from mammalian brain: Comparative peptide mapping and immunological studies with monoclonal antibodies

1986 ◽  
Vol 246 (1) ◽  
pp. 419-427 ◽  
Author(s):  
Tito Ureta ◽  
Allen D. Smith ◽  
John E. Wilson
Keyword(s):  
Monoclonal Antibodies ◽  
Peptide Mapping ◽  
Mammalian Brain

scholarly journals Monoclonal antibodies identify a group of nuclear pore complex glycoproteins.

1987 ◽  
Vol 104 (5) ◽  
pp. 1143-1156 ◽  
Author(s):  
C M Snow ◽  
A Senior ◽  
L Gerace
Keyword(s):  
Monoclonal Antibodies ◽  
Nuclear Envelope ◽  
Nuclear Pore Complex ◽  
Peptide Mapping ◽  
Polyclonal Antibodies ◽  
Integral Membrane Protein ◽  
Nucleocytoplasmic Transport ◽  
Nuclear Pore ◽  
Nuclear Surface ◽  
Punctate Pattern

Using monoclonal antibodies we identified a group of eight polypeptides of rat liver nuclear envelopes that have common epitopes. Most or all of these proteins are structurally distinct, as shown by tryptic peptide mapping and analysis with polyclonal antibodies. While these polypeptides are relatively tightly bound to nuclear membranes, only one is an integral membrane protein. The eight antigens cofractionate with the nuclear pore complex under various conditions of ionic strength and detergent. It can be seen by immunofluorescence microscopy that the monoclonal antibodies reacting with these antigens stain the nuclear surface of interphase cells in a finely punctate pattern. When the nuclear envelope is disassembled and subsequently reformed during mitosis, the proteins are reversibly dispersed throughout the cytoplasm in the form of minute foci. By EM immunogold localization on isolated nuclear envelopes, the monoclonal antibodies label exclusively the nuclear pore complex, at both its nucleoplasmic and cytoplasmic margins. Considered together, our biochemical and localization data indicate that the eight nuclear envelope polypeptides are pore complex components. As shown in the accompanying paper (Holt, G. D., C. M. Snow, A. Senior, R. S. Haltiwanger, L. Gerace, and G. W. Hart, J. Cell Biol., 104:1157-1164) these eight polypeptides contain a novel form of glycosylation, O-linked N-acetylglucosamine. The relative abundance and disposition of these O-linked glycoproteins in the pore complex are consistent with their having a role in nucleocytoplasmic transport.

scholarly journals Monoclonal antibodies against a C-terminal peptide of human brain acetylcholinesterase distinguish between erythrocyte and brain acetylcholinesterases

1996 ◽  
Vol 42 (1) ◽  
pp. 19-23 ◽  
Author(s):  
N Boschetti ◽  
U Brodbeck ◽  
S P Jensen ◽  
C Koch ◽  
B Nørgaard-Pedersen
Keyword(s):  
Monoclonal Antibodies ◽  
Human Brain ◽  
Solid Phase ◽  
Bovine Brain ◽  
Mammalian Brain ◽  
Dot Blot ◽  
Electric Eel ◽  
Terminal Amino ◽  
Dot Blot Analysis ◽  
Brain Acetylcholinesterase

Abstract Monoclonal antibodies (mAbs) were raised against a peptide of the 10 C-terminal amino acids of human brain acetylcholinesterase (AChE): H-Tyr-Ser-Lys-Gln-Asp-Arg-Cys-Ser-Asp-Leu-OH. Two positive clones (mAbs 190-1 and 190-2) were selected and tested for their ability to distinguish between mammalian brain and erythrocyte AChEs. In a solid-phase enzyme antigen immunoassay as well as by Western- and dot-blot analysis, both antibodies showed clear binding to AChE from human and bovine brain but not to AChE from erythrocytes. MAbs 190-1 and 190-2 reacted with neither AChE from electric eel nor butyrylcholinesterase from human serum. Both antibodies were used in a quantitative assay for AChE in amniotic fluids, where AChE activity could be found only in samples from open neural tube-defect pregnancies, but not in fluids from normal pregnancies or in artificially blood-contaminated samples.

scholarly journals Human proteasomes analysed with monoclonal antibodies

1995 ◽  
Vol 305 (1) ◽  
pp. 245-252 ◽  
Author(s):  
K B Hendil ◽  
P Kristensen ◽  
W Uerkvitz
Keyword(s):  
Monoclonal Antibodies ◽  
Peptide Mapping ◽  
Protein Complexes ◽  
Electrophoretic Analysis ◽  
Eukaryotic Cells ◽  
Two Dimensional Electrophoresis ◽  
Cell Extracts ◽  
Liver And Kidney ◽  
Alpha Type ◽  
Dimensional Electrophoresis

The proteasome or multicatalytic endopeptidase from eukaryotic cells consists of at least 14 subunits that fall into two families, alpha and beta. Subunit-specific monoclonal antibodies against ten different subunits of human proteasomes have been produced, together with an antibody that reacts with a motif (prosbox 1), common to alpha-type subunits. Four of the subunit-specific antibodies were able to precipitate proteasomes. The subunit composition of HeLa-cell proteasomes precipitated with these four different antibodies were identical, as judged from two-dimensional electrophoresis. One of the four antibodies was used to obtain proteasomes from cell lines (HeLa, Daudi, IMR90 and BSC-1) and human tissues (placenta, kidney, and liver). Electrophoretic analysis of these proteasomes, combined with peptide mapping of some subunits, suggests that they all contain 14 types of subunits as their major constituents. However, one subunit was present in two isoelectric isoforms in all cells examined. Two other subunits occurred in two or three isoelectric isoforms in placenta, liver and kidney, but not in the cell cultures. Extracts of human cells (HeLa, IMR90, Daudi and erythrocytes) were analysed by non-denaturing electrophoresis and immunoblotting. All of the 11 subunits detected by antibodies were present in a pair of ATP-stabilized protein complexes, presumed to be the 26 S proteinase, and in a doublet of complexes which migrated more slowly than purified proteasomes. Besides being present in proteasomes, one subunit was also found to occur in the free state in cell extracts.

Mammalian brain antigens defined by monoclonal antibodies

1984 ◽  
Vol 322 (2) ◽  
pp. 277-287 ◽  
Author(s):  
Angel L. De Blas ◽  
Rodrigo O. Kuljis ◽  
Holly M. Cherwinski
Keyword(s):  
Monoclonal Antibodies ◽  
Mammalian Brain

scholarly journals Molecular localization of human class II MT2 and MT3 determinants.

1984 ◽  
Vol 160 (2) ◽  
pp. 472-493 ◽  
Author(s):  
C K Hurley ◽  
R C Giles ◽  
G Nunez ◽  
R DeMars ◽  
L Nadler ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Amino Acid ◽  
Cell Surface ◽  
Peptide Mapping ◽  
Amino Acid Sequencing ◽  
Human Class ◽  
Panel Testing ◽  
Complement Dependent Cytotoxicity ◽  
Surface Fluorescence ◽  
D Region

The specificities of the monoclonal antibodies I-LR2 and 109d6, which recognize MT2- and MT3-like serologic determinants, respectively, have been confirmed by panel testing. In addition, the relationships of these antibodies to other monoclonal antibodies and alloantisera have been studied by means of cell surface fluorescence, complement-dependent cytotoxicity and immunoprecipitation. Using these monoclonal antibodies, molecules encoded by the HLA-D region have been isolated and characterized by amino acid sequencing and peptide mapping. By these criteria, the major populations of molecules bearing MT2- and MT3-like determinants are indistinguishable from DR molecules.

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