scholarly journals A collagenous glycoprotein found in dissociative extracts of foetal bovine nuchal ligament. Evidence for a relationship with type VI collagen

1984 ◽  
Vol 220 (2) ◽  
pp. 395-403 ◽  
Author(s):  
K R Knight ◽  
S Ayad ◽  
C A Shuttleworth ◽  
M E Grant
Keyword(s):  
Amino Acid ◽  
Ion Exchange ◽  
Amino Acid Analysis ◽  
Gel Filtration ◽  
Enzyme Digestion ◽  
Type Vi Collagen ◽  
Pepsin Digestion ◽  
Bacterial Collagenase ◽  
Collagenase Digestion ◽  
Nuchal Ligament

A collagenous glycoprotein (Mr 140000) was isolated from dissociative extracts of foetal bovine nuchal ligament and purified by a combination of ion-exchange and gel-filtration chromatography. This glycoprotein (designated MFPI) exists as a large-Mr disulphide-bonded aggregate in the absence of a reducing agent. The purified glycoprotein was shown to contain about 6% (w/w) carbohydrate, mostly as galactose, glucose and mannose. Amino acid analysis showed the presence of hydroxyproline and hydroxylysine, indicative of its collagenous nature. The collagenous nature of this glycoprotein was further investigated by enzyme digestion. Pepsin digestion produced three major fragments, which were identical with peptides of type VI collagen. Bacterial-collagenase digestion of the unreduced glycoprotein also produced several discrete peptides. However, reduction of the glycoprotein before bacterial-collagenase digestion resulted in the degradation of these discrete peptides. Glycoprotein MFPI extracted in dissociative conditions appears to be a larger-Mr form of type VI collagen, believed to originate from microfibrillar components in the intact tissue.

Isolation and ultrastructural analysis of microfibrillar structures from foetal bovine elastic tissues. Relative abundance and supramolecular architecture of type VI collagen assemblies and fibrillin

1991 ◽  
Vol 99 (4) ◽  
pp. 797-807
Author(s):  
C.M. Kielty ◽  
C. Cummings ◽  
S.P. Whittaker ◽  
C.A. Shuttleworth ◽  
M.E. Grant
Keyword(s):  
Relative Abundance ◽  
Gel Filtration ◽  
Supramolecular Architecture ◽  
Collagen Vi ◽  
Type Vi Collagen ◽  
Bacterial Collagenase ◽  
New Information ◽  
Sds Page ◽  
Complex Polymers ◽  
Nuchal Ligament

Extensive intact assemblies of matrix macromolecules have been solubilized from foetal calf skin, nuchal ligament and aorta by a new procedure that includes bacterial collagenase digestion under non-reducing, non-denaturing conditions and gel filtration chromatography. Type VI collagen was identified as the major microfibrillar element of these tissues by SDS-PAGE analysis and Western blotting. Rotary shadowing electron microscopy of these preparations revealed by far the most abundant and extensive arrays of intact collagen VI microfibrils isolated to date. The distinct microfibrillar species, fibrillin, which was identified on the basis of its periodicity and morphology, was also solubilized in abundance by this protocol. Analysis of these complex polymers has generated new information on their supramolecular architecture and relative abundance in these tissues. The protocol also demonstrates that the release of intact collagen VI microfibrils from these tissues is largely dependent on the removal of the major collagen fibrils.

scholarly journals Isolation and a partial amino acid sequence of insulin from the islet tissue of cod (Gadus callarias)

1968 ◽  
Vol 106 (2) ◽  
pp. 531-541 ◽  
Author(s):  
P T Grant ◽  
K. B. M. Reid
Keyword(s):  
Amino Acid ◽  
Ion Exchange ◽  
Gel Filtration ◽  
Bovine Insulin ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Performic Acid ◽  
Amino Acid Residues ◽  
Islet Tissue ◽  
A Chain

1. Insulin has been isolated by gel filtration and ion-exchange chromatography from extracts of the discrete islet tissue of cod. The final preparation yielded a single band on electrophoresis at two pH values. The biological potency was 11·5 international units/mg. in mouse-convulsion and other assay procedures. 2. Glycine and methionine were shown to be the N-terminal amino acids of the A and B chains respectively. An estimate of the molecular weight together with amino acid analyses indicated that cod insulin, like the bovine hormone, consists of 51 amino acid residues. In contrast, the amino acid composition differs markedly from bovine insulin. 3. Oxidation of insulin with performic acid yielded the A and B peptide chains, which were separated by ion-exchange chromatography. Sequence studies on smaller peptides isolated from enzymic digests or from dilute acetic acid hydrolysates of the two chains have established the sequential order of 14 of the 21 amino acid residues of the A chain and 25 of the 30 amino acid residues of the B chain.

scholarly journals Purification and characterization of human neuropeptide Y from adrenal-medullary phaeochromocytoma tissue

1984 ◽  
Vol 219 (3) ◽  
pp. 699-706 ◽  
Author(s):  
R Corder ◽  
P C Emson ◽  
P J Lowry
Keyword(s):  
Amino Acid ◽  
Neuropeptide Y ◽  
Amino Acid Analysis ◽  
Gel Filtration ◽  
Exchange Chromatography ◽  
Carboxypeptidase Y ◽  
Purification And Characterization ◽  
Reverse Phase ◽  
Tryptic Fragment

Human neuropeptide Y was isolated from acid extracts of adrenal-medullary phaeochromocytoma tissue. After (NH4)2SO4 fractionation, the neuropeptide Y-like immunoreactivity was purified from the resolubilized 80%-saturation-(NH4)2SO4 peptide-rich precipitate, by gel filtration, cation-exchange chromatography and reverse-phase high-pressure liquid chromatography. Amino acid analysis of the peptide revealed a composition almost identical with that of the pig peptide, the exception being the loss of one leucine residue and its replacement with methionine. Tryptic digestion of the peptide and subsequent amino acid analysis of the fragments further confirmed the identity of the peptide. Carboxypeptidase Y digestion of the (1-19)-peptide tryptic fragment has shown the methionine to be located at position 17 in human neuropeptide Y.

scholarly journals The isolation of three neurophysins from porcine posterior pituitary lobes

1970 ◽  
Vol 116 (5) ◽  
pp. 899-909 ◽  
Author(s):  
L. O. Uttenthal ◽  
D. B. Hope
Keyword(s):  
Molecular Weight ◽  
Amino Acid ◽  
Amino Acid Analysis ◽  
Gel Filtration ◽  
Biological Significance ◽  
Starch Gel Electrophoresis ◽  
Posterior Pituitary ◽  
Fresh Tissue ◽  
Molecular Weights ◽  
Molecular Dimensions

1. Three neurophysins, proteins that bind the polypeptide hormones oxytocin and vasopressin, have been isolated from acetone-dried porcine posterior pituitary lobes. The proteins have been named porcine neurophysins-I, -II and -III in order of their electrophoretic mobilities at pH8.1. 2. Electrophoretic comparison of the purified proteins, which are homogeneous on starch-gel electrophoresis, with the soluble proteins of fresh porcine posterior pituitary lobes extracted in 0.1m-HCl and in buffer pH8.1 suggests that the isolated proteins are native to the fresh tissue. 3. Neurophysins-I and -II are present in similar amounts in the tissue, whereas neurophysin-III is present only in small quantities. Acetone-dried tissue also contains traces of other hormone-binding neurophysin components. 4. All the neurophysins can bind both oxytocin and [8-lysine]-vasopressin. 5. The apparent molecular weights of the neurophysins increase with increasing protein concentration as measured by equilibrium sedimentation in the ultracentrifuge. 6. Neurophysins-I and -III are of similar molecular dimensions, contain one residue of methionine per molecule and lack histidine. The minimum molecular weight of neurophysin-I obtained by amino acid analysis is 9360. Neurophysin-II is of larger molecular dimensions than neurophysins-I and -III and can be separated from these by gel filtration on Sephadex G-75. It contains no histidine or methionine, and its minimum molecular weight has been estimated as 14020 by amino acid analysis. 7. Each of the three neurophysins possesses N-terminal alanine. 8. The possible biological significance of the existence of several neurophysins within one species is discussed.

scholarly journals Purification and Characterization of Catalase from Marine BacteriumAcinetobactersp. YS0810

2014 ◽  
Vol 2014 ◽  
pp. 1-7 ◽  
Author(s):  
Xinhua Fu ◽  
Wei Wang ◽  
Jianhua Hao ◽  
Xianglin Zhu ◽  
Mi Sun
Keyword(s):  
Amino Acid ◽  
Ion Exchange ◽  
Gel Filtration ◽  
Protoporphyrin Ix ◽  
Hydroxylamine Hydrochloride ◽  
Sodium Dithionite ◽  
Flight Mass Spectrometry ◽  
Alkali Stability ◽  
High Alkali ◽  
High Degree

The catalase from marine bacteriumAcinetobactersp. YS0810 (YS0810CAT) was purified and characterized. Consecutive steps were used to achieve the purified enzyme as follows: ethanol precipitation, DEAE Sepharose ion exchange, Superdex 200 gel filtration, and Resource Q ion exchange. The active enzyme consisted of four identical subunits of 57.256 kDa. It showed a Soret peak at 405 nm, indicating the presence of iron protoporphyrin IX. The catalase was not apparently reduced by sodium dithionite but was inhibited by 3-amino-1,2,4-triazole, hydroxylamine hydrochloride, and sodium azide. Peroxidase-like activity was not found with the substrate o-phenylenediamine. So the catalase was determined to be a monofunctional catalase. N-terminal amino acid of the catalase analysis gave the sequence SQDPKKCPVTHLTTE, which showed high degree of homology with those of known catalases from bacteria. The analysis of amino acid sequence of the purified catalase by matrix-assisted laser desorption ionization time-of-flight mass spectrometry showed that it was a new catalase, in spite of its high homology with those of known catalases from other bacteria. The catalase showed high alkali stability and thermostability.

Ion-exchange chromatography for physiological fluid amino acid analysis

1991 ◽  
Vol 2 (12) ◽  
pp. 671-679 ◽  
Author(s):  
Lean Teik Ng ◽  
David Y. Wong ◽  
Thomas Francis ◽  
G. Harvey Anderson
Keyword(s):  
Amino Acid ◽  
Ion Exchange ◽  
Amino Acid Analysis ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Physiological Fluid

scholarly journals The Amino Acid Sequence of the Large Plakalbumin Peptide and the C-Terminal Sequence of Ovalbumin

1971 ◽  
Vol 24 (3) ◽  
pp. 525 ◽  
Author(s):  
EOP Thompson ◽  
RW Slelgh ◽  
MB Smith
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Gel Filtration ◽  
Enzyme Digestion ◽  
Terminal Portion ◽  
Terminal Sequence ◽  
Hydrophobic Areas

The amino acid sequence of a 33-residue peptide isolated from plakalbumin by gel filtration in 6M urea at pH 3 and derived from the C-terminal portion of ovalbumin has been determined. Enzyme digestion of the hydrophobic areas by thermolysin, papain, ana subtilisin BPN' gave peptides with overlapping sequences. The peptides were fractionated by a combination of paper ionophoresis and chromatography and their sequences determined by the dansyl-Edman technique.

scholarly journals Studies on Marsupial Protein. II. Amino Acid Sequence of the -Chain of Haemoglobin from the Grey Kangaroo, Macropus Giganteus

1969 ◽  
Vol 22 (6) ◽  
pp. 1437 ◽  
Author(s):  
GM Air ◽  
EOP Thompson
Keyword(s):  
Amino Acid ◽  
Ion Exchange ◽  
Amino Acid Sequence ◽  
Gel Filtration ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Amino Acid Sequences ◽  
Tryptic Digestion ◽  
Grey Kangaroo

The amino acid sequence of the jS-chain of haemoglobin from M. giganteus has been determined. The soluble peptides formed by tryptic digestion were isolated by gel filtration, ion-exchange chromatography, and paper ionophoresis, and amino acid sequences determined by the "dansyl"-Edman procedure. Special procedures were necessary for three peptides which were insoluble.

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