Primary structure and tissue-specific expression of blue crab (Callinectes sapidus) metallothionein isoforms

In aquatic animals, synthesis of the metal-binding protein metallothionein (MT) can be induced through exposure to elevated levels of metals in food or water. Whether the different routes of exposure lead to expression of different metallothionein isoforms in different tissues in unknown. In this study we examined the induction of metallothionein isoforms in the hepatopancreas and gills of the blue crab Callinectes sapidus. When blue crabs are exposed to cadmium in their diet, the metal accumulates in the hepatopancreas. Size-exclusion and anion-exchange chromatography show the presence of five low-molecular-mass cadmium-binding proteins. All of the observed cadmium-binding proteins belong to the class I MT family. They are designated as MT-Ia, MT-Ib, MT-Ic, MT-IIa and MT-IIb. All purified proteins run as single peaks upon rechromatography on anion-exchange HPLC, except for MT-Ic, which segregates into two peaks corresponding to MT-Ia and MT-Ic. The amino acid sequence of MT-Ia and MT-Ic is identical. MT-Ib differs from MT-Ia and MT-Ic only in having an extra N-terminal methionine. The 18 cysteine residues in MT-Ia and MT-IIa occur in identical positions; however, of the remaining 40 amino acids, 15 are found to be different. MT-IIb is identical with MT-IIa, except for an extra methionine residue at its N-terminal position. It appears therefore that, of the five observed CdMTs, only two are the products of distinct genes. CdMT-Ia and -IIa are posttranslationally modified forms of Ib and IIb, respectively, and CdMT-Ia and -Ic appear to be conformational isomers. Cadmium-induced expression of the two genes is tissue-specific. When crabs are exposed to cadmium in water, the metal accumulates in the gills, where it is bound to MT-II. MT-I is virtually absent.

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Cadmium-binding proteins of rat testes. Characterization of a low-molecular-mass protein that lacks identity with metallothionein

Biochemical Journal ◽

10.1042/bj2200811 ◽

1984 ◽

Vol 220 (3) ◽

pp. 811-818 ◽

Cited By ~ 50

Author(s):

M P Waalkes ◽

S B Chernoff ◽

C D Klaassen

Keyword(s):

Metal Binding ◽

Gel Electrophoresis ◽

Binding Proteins ◽

Polyacrylamide Gel Electrophoresis ◽

Sodium Dodecyl ◽

Gel Filtration ◽

Anion Exchange Chromatography ◽

Exchange Chromatography ◽

Polyacrylamide Gel ◽

Or Groups

Cadmium-binding proteins in the cytosol of testes from untreated rats were separated by Sephadex G-75 gel filtration. Three major testicular metal-binding proteins (TMBP), or groups of proteins, with relative elution volumes of approx. 1.0 (TMBP-1), 1.7 (TMBP-2) and 2.4 (TMBP-3) were separated. Elution of Zn-binding proteins exhibited a similar pattern. TMBP-3 has previously been thought to be metallothionein (MT), and hence this protein was further characterized and compared with hepatic MT isolated from Cd-treated rats. Estimation of Mr by gel filtration indicated a slight difference between MT (Mr 10000) and TMBP-3 (Mr 8000). Two major forms of MT (MT-I and MT-II) and TMBP-3 (TMBP-3 form I and TMBP-3 form II) were obtained after DEAE-Sephadex A-25 anion-exchange chromatography, with the corresponding subfractions being eluted at similar conductances. Non-denaturing polyacrylamide-gel electrophoresis on 7% acrylamide gels indicated that the subfractions of TMBP-3 had similar mobilities to those of the corresponding subfractions of MT. However, SDS (sodium dodecyl sulphate)/12% (w/v)-polyacrylamide-gel electrophoresis resulted in marked differences in migration of the two corresponding forms of MT and TMBP-3. Co-electrophoresis of MT-II and TMBP-3 form II by SDS/polyacrylamide-gel electrophoresis revealed two distinct proteins. Amino acid analysis indicated much lower content of cysteine in the testicular than in the hepatic proteins. TMBP-3 also contained significant amounts of tyrosine, phenylalanine and histidine, whereas MT did not. U.v.-spectral analysis of TMBP-3 showed a much lower A250/A280 ratio than for MT. Thus this major metal-binding protein in testes, which has been assumed to be MT is, in fact, a quite different protein.

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Tissue-Specific Expression of Messenger Ribonucleic Acids for Insulin-Like Growth Factors and Insulin-Like Growth Factor-Binding Proteins during Perinatal Development of the Rat Uterus1

Biology of Reproduction ◽

10.1095/biolreprod60.5.1172 ◽

1999 ◽

Vol 60 (5) ◽

pp. 1172-1182 ◽

Cited By ~ 24

Author(s):

Y. Gu ◽

W.S. Branham ◽

D.M. Sheehan ◽

P.J. Webb ◽

C.L. Moland ◽

...

Keyword(s):

Growth Factors ◽

Growth Factor ◽

Binding Proteins ◽

Specific Expression ◽

Tissue Specific ◽

Ribonucleic Acids ◽

Perinatal Development ◽

Factor Binding ◽

Tissue Specific Expression ◽

Insulin Like Growth Factors

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Analysis of the substituent distribution along the chain of water-soluble methyl cellulose by combination of enzymatic and chemical methods

Holzforschung ◽

10.1515/hf.2004.013 ◽

2004 ◽

Vol 58 (1) ◽

pp. 97-104 ◽

Cited By ~ 10

Author(s):

B. Saake ◽

S. Lebioda ◽

J. Puls

Keyword(s):

Anion Exchange ◽

Size Exclusion Chromatography ◽

Methyl Cellulose ◽

Anion Exchange Chromatography ◽

Exchange Chromatography ◽

Degree Of Substitution ◽

Water Soluble ◽

Size Exclusion ◽

Exclusion Chromatography ◽

C Nmr

Abstract Four methyl cellulose samples in the degree of substitution range from 0.5 to 2.0 were characterised by combination of different analytical methods. Samples were analysed regarding their partial degree of substitution by hydrolysis and anion exchange chromatography with pulsed amperometric detection. For calibration of the chromatographic system, standard substances were isolated by preparative HPLC and their structure was confirmed by 13C-NMR spectroscopy. For two methyl cellulose samples per-acetylation and 13C-NMR with inverse gated decoupling was carried out for comparison with the chromatographic analysis. Endoglucanase fragmentation of methyl celluloses was performed and water-soluble and insoluble fractions were analysed separately. A preparative size exclusion chromatography system for enzymatic-degraded water-soluble methyl cellulose was developed and the molar masses of the individual fractions were examined by analytical size exclusion chromatography. By combination of endoglucanase fragmentation, preparative chromatography, hydrolysis and anion exchange chromatography an approach for the analysis of the substitutent distribution along the polymeric chain of water-soluble methyl cellulose could be established.

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Purification of cell culture-derived human influenza A virus by size-exclusion and anion-exchange chromatography

Biotechnology and Bioengineering ◽

10.1002/bit.21109 ◽

2007 ◽

Vol 96 (5) ◽

pp. 932-944 ◽

Cited By ~ 66

Author(s):

Bernd Kalbfuss ◽

Michael Wolff ◽

Robert Morenweiser ◽

Udo Reichl

Keyword(s):

Cell Culture ◽

Anion Exchange ◽

Influenza A Virus ◽

Influenza A ◽

Anion Exchange Chromatography ◽

Exchange Chromatography ◽

Size Exclusion ◽

Human Influenza

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Peer Review #2 of "Sex- and tissue-specific expression of odorant-binding proteins and chemosensory proteins in adults of the scarab beetle Hylamorpha elegans (Burmeister) (Coleoptera: Scarabaeidae) (v0.2)"

10.7287/peerj.7054v0.2/reviews/2 ◽

2019 ◽

Keyword(s):

Peer Review ◽

Binding Proteins ◽

Scarab Beetle ◽

Specific Expression ◽

Tissue Specific ◽

Odorant Binding Proteins ◽

Chemosensory Proteins ◽

Tissue Specific Expression ◽

Odorant Binding

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Isolation and Characterization of the Vascular-specific 22-kDa Zn-binding Protein

HortScience ◽

10.21273/hortsci.32.3.475d ◽

1997 ◽

Vol 32 (3) ◽

pp. 475D-475

Author(s):

Kathryn C. Taylor ◽

Danielle R. Elli

Keyword(s):

Amino Acid ◽

Metal Binding ◽

Binding Protein ◽

Xylem Sap ◽

Exchange Chromatography ◽

Size Exclusion ◽

Ancestral Gene ◽

Rangpur Lime ◽

Rough Lemon ◽

Isolation And Characterization

A 22-kDa Zn-binding protein (ZBP) was isolated from the phloem tissue and evacuated xylem sap of `Valencia' sweet orange [Citrus sinensis (L.) Osbeck] on rough lemon [C. jambhiri (L.)], as well as Valencia on Rangpur lime [Citrus limonia Osbeck]. Phloem and xylem Zn was associated with the 22 kDa ZBP. The Mr value of this ZBP was estimated to be 19,500 by size exclusion chromatography and 22,800 by SDS-PAGE. This protein was isolated with an isoelectric point of 7.5. Ion exchange chromatography demonstrated that 22-kDa ZBP was highly anionic, requiring 0.43 M NaCl for elution from QAE Sepharose. The 22-kDa ZBP appears unique to citrus, having no cross reaction with protein from several tissues from a range of plant species. Accumulation decreased under Zn-deficient conditions, was enhanced by osmotic stress, and the protein completely disappeared with wounding. Amino acid composition demonstrated that the protein was rich in aspartate, and glutamate; and contained 6 cysteine, and 4 histidine residues. These amino acids may be involved in metal binding. N-terminal amino acid sequencing demonstrated that the 22-kDa ZBP had identity with sporamin A&B precursors, Kunitz-type trypsin inhibitors, and miraculin. It is suggested that the genes that encode these proteins are derived from a common ancestral gene.

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Peer Review #2 of "Sex- and tissue-specific expression of odorant-binding proteins and chemosensory proteins in adults of the scarab beetle Hylamorpha elegans (Burmeister) (Coleoptera: Scarabaeidae) (v0.1)"

10.7287/peerj.7054v0.1/reviews/2 ◽

2019 ◽

Keyword(s):

Peer Review ◽

Binding Proteins ◽

Scarab Beetle ◽

Specific Expression ◽

Tissue Specific ◽

Odorant Binding Proteins ◽

Chemosensory Proteins ◽

Tissue Specific Expression ◽

Odorant Binding

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Purification and N-terminal amino acid sequence of a chondroitin sulphate/dermatan sulphate proteoglycan isolated from intima/media preparations of human aorta

Biochemical Journal ◽

10.1042/bj2740415 ◽

1991 ◽

Vol 274 (2) ◽

pp. 415-420 ◽

Cited By ~ 20

Author(s):

G Stöcker ◽

H E Meyer ◽

C Wagener ◽

H Greiling

Keyword(s):

Anion Exchange ◽

Core Protein ◽

Chondroitin Sulphate ◽

Exchange Chromatography ◽

Size Exclusion ◽

Human Aorta ◽

Dermatan Sulphate ◽

Western Blots ◽

Terminal Amino ◽

Differential Digestion

A proteoglycan (PG) was purified to homogeneity from intima/media preparations of human aorta specimens by the following chromatographic steps: Sepharose Q anion exchange, Sepharose CL-4B size exclusion, hydroxyapatite, MonoQ anion exchange and TSK G 4000 SW size exclusion. The purity of the preparation was established by SDS/PAGE using direct staining by silver or Dimethylmethylene Blue, as well as by Western blots of biotin-labelled samples. The electrophoretic mobility of the native PG was less than that of a 200,000-Mr standard protein. After treatment with chondroitin sulphate lyase ABC, a core protein of Mr 15,000 was revealed. The Mr of the glycosaminoglycan (GAG) peptides was less than 24,000, by comparison with a keratan sulphate peptide. The composition of the GAG chains was determined by differential digestion of the PG by chondroitin sulphate lyases AC/ABC or chondroitin sulphate lyase AC alone followed by anion-exchange chromatography of the resulting disaccharides. The GAG chains are composed of approximately one-third of dermatan sulphate and two-thirds chondroitin sulphate disaccharide units. The sequence of the 20 N-terminal amino acids is identical with the sequence previously reported for PG I isolated from human developing bone [Fisher, Termine & Young (1989) J. Biol. Chem. 264, 4571-4576]. The assignment of glycosylation sites to the serine residues in positions 5 and 10 was confirmed. The findings indicate that the chondroitin sulphate/dermatan sulphate PG is a major PG in intima/media preparations of human aorta and represents a biglycan-type PG.

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