scholarly journals Biochemical analysis of actin in crane-fly gonial cells: evidence for actin in spermatocytes and spermatids--but not sperm.

1980 ◽  
Vol 86 (1) ◽  
pp. 315-325 ◽  
Author(s):  
A R Strauch ◽  
E J Luna ◽  
J R LaFountain
Keyword(s):  
Molecular Weight ◽  
Comparative Analysis ◽  
Gel Electrophoresis ◽  
Biochemical Analysis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Cell Types ◽  
Two Dimensional ◽  
Peptide Analysis ◽  
General Applicability ◽  
Peptide Maps

A biochemical assay employing DNase-I affinity chromatography, two-dimensional peptide analysis and SDS polyacrylamide gel electrophoresis was used to isolate, identify, and assess the amount of actin from gonial cells of the crane fly, Nephrotoma suturalis. Based on the analysis of cell homogenates under conditions in which all cellular actin is converted to the monomeric DNase-binding form, actin comprises approximately 1% of the total protein in homogenates of spermatocytes and spermatids. SDS gel analysis of mature sperm reveals no polypeptides with a molecular weight similar to that of actin. Under conditions that preserve native supramolecular states of actin, approximately 80% of the spermatocyte actin is in a sedimentable form whereas only approximately 30% of the spermatid actin is sedimentable. These differences could be meaningful with regard to strutural changes that occur during spermiogenesis. A comparative analysis of two-dimensional peptide maps of several radioiodinated actins reveals similarities among spermatocyte, spermatid, and human erythrocyte actins. The results suggest the general applicability of this approach to other cell types that contain limited amounts of actin.

Variation in goat fibre protein

1989 ◽  
Vol 40 (3) ◽  
pp. 675 ◽  
Author(s):  
DJ Tucker ◽  
AHF Hudson ◽  
A Laudani ◽  
RC Marshall ◽  
DE Rivett
Keyword(s):  
Molecular Weight ◽  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Apparent Molecular Weight ◽  
Wool Fibre ◽  
Two Dimensional ◽  
Protein Patterns ◽  
Cashmere Goats

The proteins from a range of cashmere, mohair, angoratcashmere crossbred and wool fibre samples were extracted at pH 8 with 8 M urea containing dithiothreitol, and were then radiolabelled by S-carboxymethylation using iodo(2-14C) acetate. The proteins from each sample were examined by two dimensional polyacrylamide gel electrophoresis in which the separation in the first dimension was according to charge at pH 8.9 and in the second dimension according to apparent molecular weight in the presence of sodium dodecyl sulfate. After electrophoresis the proteins were detected by fluorography. Protein differences in keratin samples from some individual goats existed, although the overall protein patterns were similar. None of the differences were consistent with any one goat fibre type. The protein patterns obtained for fibre samples from individual cashmere goats showed some differences when compared to those found for commercial blends from the same country of origin, indicating that blending can mask any animal-to-animal variation. While the electrophoretic technique does not unequivocally distinguish between cashmere, mohair and angora/cashmere crossbred fibres it does differentiate between wool and goat fibres.

Structure of the O-chain of the lipopolysaccharide of Haemophilus pleuropneumoniae serotype 1

1986 ◽  
Vol 64 (12) ◽  
pp. 1317-1325 ◽  
Author(s):  
Eleonora Altman ◽  
Jean-Robert Brisson ◽  
Malcolm B. Perry
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Lipid A ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Two Dimensional ◽  
Magnetic Resonance Studies ◽  
Serotype 1 ◽  
Structure Formula ◽  
Phenol Water

By phenol-water extraction an aqueous-phase soluble cellular lipopolysaccharide was isolated from Haemophilus pleuropneumoniae serotype 1. It was shown by sodium dodecyl sulfate – polyacrylamide gel electrophoresis, hydrolysis, methylation, and both one- and two-dimensional 1H and 13C nuclear magnetic resonance studies to be an S-type lipopolysaccharide, which could be cleaved to yield a lipid A and an O-chain polysaccharide identified as a high molecular weight branched polymer of a tetrasaccharide repeating unit having the structure:[Formula: see text]

scholarly journals The distinguishing characteristics of the plasma membrane are its exposed proteins

1975 ◽  
Vol 147 (2) ◽  
pp. 373-376 ◽  
Author(s):  
R E Gates ◽  
D R Phillips ◽  
M Morrison
Keyword(s):  
Molecular Weight ◽  
Plasma Membrane ◽  
Gel Electrophoresis ◽  
Human Lymphocytes ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Cell Types ◽  
Distinguishing Characteristic ◽  
Probe System ◽  
Normal Human

The exposed proteins of the plasma membrane of normal human lymphocytes and platelets were labelled by using the lactoperoxidase macromolecular probe system. The labelled components were separated into molecular-weight classes by sodium dodecyl sulphate--polyacrylamide-gel electrophoresis. In contrast with the report by Tanner et al. (1974), a comparison of the two cell types showed that the major labelled components in both cell types were glycoproteins and were not identical. It is concluded that the exposed proteins are probably the most distinguishing characteristic of the plasma membrane of differentiated cell types.

scholarly journals Identification of the platelet-specific alloantigen, Naka, on platelet membrane glycoprotein IV

Blood ◽  
1990 ◽  
Vol 75 (3) ◽  
pp. 684-687 ◽  
Author(s):  
Y Tomiyama ◽  
H Take ◽  
H Ikeda ◽  
T Mitani ◽  
T Furubayashi ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Platelet Membrane ◽  
Membrane Glycoprotein ◽  
Two Dimensional ◽  
Definitive Evidence ◽  
Sds Page ◽  
Platelet Transfusions

We describe the membrane localization of a new platelet-specific alloantigen, designated Naka, that is involved in refractoriness to HLA- matched platelet transfusions. By indirect immunoprecipitation, anti- Naka antibody precipitated a single, radiolabeled platelet membrane protein with a molecular weight (mol wt) of 91 Kd from Naka-positive platelets. When radiolabeled Naka-negative platelets were used as a source of target antigens, no radiolabeled proteins were precipitated. The analyses using nonreduced-reduced two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and using rabbit antiglycoprotein (GP)IV demonstrated that this protein corresponds to GPIV (alternatively GPIIIb). Furthermore, in dot immunobinding, anti- Naka antibody bound to purified GPIV. Our results provide definitive evidence that the Naka alloantigen is carried on GPIV. These results also demonstrate that, on occasion, antibodies against GPIV may play an important role in refractoriness to platelet transfusions.

scholarly journals Mammalian chromatin substructure studies with the calcium–magnesium endonuclease and two-dimensional polyacrylamide-gel electrophoresis

1974 ◽  
Vol 143 (1) ◽  
pp. 67-72 ◽  
Author(s):  
Leigh A. Burgoyne ◽  
Dean R. Hewish ◽  
Jennifer Mobbs
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Nuclear Dna ◽  
Polyacrylamide Gel Electrophoresis ◽  
Dna Fragments ◽  
Two Dimensional ◽  
Digestion Time ◽  
Calcium Magnesium ◽  
Liver Chromatin ◽  
Time Courses

The basic regularity of chromatin substructure that has been reported in rat liver chromatin (Hewish & Burgoyne, 1973b) was also detected in mouse chromatin. The regular series of DNA fragments produced by the action of Ca–Mg endonuclease on rat chromatin were studied further. The smallest single-stranded class has a molecular weight of approx. 45000–63000 and the smallest double-stranded class has a molecular weight of approx. 120000–150000. Studies of the substructure of the DNA fragments produced by the Ca–Mg endonuclease have shown that the regular series of double-stranded fragments have regular series of single-stranded fragments within them. It was concluded that the regular series of double-stranded fragments was probably a consequence of the regular series of single-stranded fragments. Digestion time-courses are presented for mouse and rat nuclear DNA.

scholarly journals Additional glycoprotein defects in Bernard-Soulier's syndrome: confirmation of genetic basis by parental analysis

Blood ◽  
1983 ◽  
Vol 62 (4) ◽  
pp. 800-807 ◽  
Author(s):  
MC Berndt ◽  
C Gregory ◽  
BH Chong ◽  
H Zola ◽  
PA Castaldi
Keyword(s):  
Molecular Weight ◽  
Gel Electrophoresis ◽  
Membrane Surface ◽  
Polyacrylamide Gel Electrophoresis ◽  
Polyacrylamide Gel ◽  
Two Dimensional ◽  
Membrane Glycoproteins ◽  
Glycoprotein Ib ◽  
Membrane Complex ◽  
Platelet Protein

Abstract The glycoprotein profile of Bernard-Soulier platelets was examined by labeling washed platelets with periodate 3H-sodium borohydride, a procedure that labels greater than 30 glycoproteins on the membrane surface of normal platelets. Three Bernard-Soulier patients were studied; two were siblings and the third was unrelated. The platelet protein and glycoprotein profiles were evaluated under nonreduced and reduced conditions using 5%-15% exponential SDS-polyacrylamide gel electrophoresis. The two siblings completely lacked glycoprotein Ib (GPIb). The unrelated patient had congruent to 7% of the normal level. This was confirmed by two-dimensional nonreduced-reduced SDS- polyacrylamide gel electrophoresis, a procedure that allows clear separation of the disulfide-linked subunits of GPIb, GPIb alpha (mol wt 145,000), and GPIb beta (mol wt 25,000) from other membrane glycoproteins. On one-dimensional analysis, Bernard-Soulier's syndrome (BSS) platelets also lacked the peripheral membrane glycoprotein, GPV (mol wt 82,000) and a low molecular weight glycoprotein, GPIX, (nonreduced or reduced, mol wt congruent to 22,000). The two- dimensional gel system also revealed the absence of a minor glycoprotein with a molecular weight of congruent to 100,000 (GP 100). Quantitation of these proteins solubilized from electrophoretograms showed that the siblings' parents had congruent to 50% levels of GPIb, GPIX, and GP 100. A monoclonal antibody against glycoprotein Ib, FMC 25, was negative by immunofluorescence against Bernard-Soulier platelets and immuneprecipitated both GP Ib and GPIX from Triton X100 solubilized, labeled platelets. The combined results suggest that the apparent genetic absence of multiple proteins in Bernard-Soulier platelets is due, in part, to the presence in normal platelets of a tight membrane complex between glycoprotein Ib and at least one of the other absent glycoproteins.

Irreversible heat denaturation of bovine α-lactalbumin

1986 ◽  
Vol 53 (2) ◽  
pp. 249-258 ◽  
Author(s):  
Lesley C. Chaplin ◽  
Richard L. J. Lyster
Keyword(s):  
Molecular Weight ◽  
Sodium Dodecyl Sulphate ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Heat Denaturation ◽  
Two Dimensional ◽  
Native Protein ◽  
Denatured Protein ◽  
Hydrolysis Of

SUMMARYThe irreversible heat denaturation of α-lactalbumin (α-la) in 0·1 M-phosphate, pH 7·0, at 100 °C was studied using polyacrylamide-gel electrophoresis (PAGE). PAGE revealed two groups of bands, one moving faster than native α-la and one slower, in addition to some denatured protein which remained at the origin and some residual native α-la. The faster group had unchanged molecular weight, but an increase in charge, partly due to hydrolysis of glutamine and asparagine residues. The slower group was shown by two-dimensional sodium dodecyl sulphate-PAGE to be oligomers of denatured α-la; formation of the smaller oligomers preceded the larger ones. The oligomers reverted to monomers in the presence of dithiothreitol, showing that they were disulphide-linked aggregates of denatured α-la. Immuno-blots of the gels showed that both fast and slow groups of bands had irreversibly lost most of the antigenicity of the native protein.

Sign in / Sign up

Export Citation Format

Share Document