Separation of Transcobalamins on Small DEAE-cellulose Columns

Gizis, Evangelos; Meyer, Leo M.

doi:10.3181/00379727-140-36451

Cited by 8 publications

(2 citation statements)

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“…DEAE-cellulose was prepared according to Gizis & Meyer (1972). Concentrated fractions (TCL and TCS), obtained from either small (a) or large (b) Sephadex columns, were applied on DEAE-cellulose columns (5 ml pipette plugged with glass wool) and elution performed with step-wise gradient of 5 sodium phosphate buffers, as reported by Kumar et a1 (1974).…”

Section: E a E-cellulose Chromatography On 'Small Columns'mentioning

confidence: 99%

Comparison of Biochemical Properties of Human Serum B₁₂ Binders Derived from Sephadex and DEAE‐Cellulose Chromatography

Kumar

Meyer

Gams

1977

Scandinavian Journal of Haematology

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Vitamin Biz binding proteins were separated into 2 peaks corresponding to small (TCS) and large (TCL) molecular weight fractions by gel-filtration on Sephadex G-200 using 0.005 M-sodium phosphate buffer, pH 7.4 containing 1 M-NaCI. Each peak, after dialysis and concentration, was chromatographed on DEAE-cellulose. 3 peaks of radioactivity were recovered from both TCS and TCL. Peaks from TCS had an apparent mol. wt. of about 40,000 and thme from TCL about 11O,OOO, as determined by gel-filtration on Sephadex G-200. On electrophoresis, peaks eluted with 0.06 Mphosphate buffer, pH 5.85 from both TCS and TCL moved as ,@-globulins; those eluted with 0.1 M buffer, pH 5.8 between pand a2-globulin and those eluted with 0.25 M buffer, pH 5.4 between q -and a2-globulin. In our assay system, TCS d e livered 57CoB1~ to L-1210 leukaemic lymphoblasts while TCL had no such activity. Of the 6 binders from DEAE-cellulose, only peaks eluted with 0.06 M and 0.1 M buffers from TCS delivered labelled Biz to these cells. Antisera prepared against TC I1 BIZ reacted only with TC I1 BIZ from serum, TCS, and fractions D and E obtained from TCS which were eluted with 0.06 and 0.1 M buffers, respectively.

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Section: E a E-cellulose Chromatography On 'Small Columns'mentioning

confidence: 99%

Comparison of Biochemical Properties of Human Serum B₁₂ Binders Derived from Sephadex and DEAE‐Cellulose Chromatography

Kumar

Meyer

Gams

1977

Scandinavian Journal of Haematology

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“…Transcobalamin I (TCI), a glyco protein [1] which probably originates from neutrophil granulocytes [21] and their precursors [24], is antigenically similar to the 'R' binders pre sent in tissue fluids [10,11,20]. By contrast, transcobalamin II (TCII) is not a glycoprotein [2], is synthesised in the liver [6,7,23], and is an tigenically different from the 'R' binders [11,22], A third binder, termed transcobalamin III (TCIII), has recently been described in nor mal human serum [4,12,13]. This binder has the same molecular rad ius and antigenic characteristics as TCI [8] and may represent a fraction resulting from a partial separation of the 3 components of TCI with dif ferent iso-electric points [17].…”

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Role of Transcobalamins I, II and III in the Transfer of Vitamin B<sub>12</sub> to Human Bone Marrow Cells <i>in vitro</i>

Wickramasinghe¹,

England²,

Saunders³

et al. 1975

Acta Haematol

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A study of the uptake of transcobalamin-bound ⁵⁷Co-cyanocobalamin by suspensions of human bone marrow cells has indicated that these cells can take up vitamin B₁₂ from all 3 transcobalamins (I, II and III). Similar transport processes were involved in the uptake from the 3 transcobalamins; uptake was dependent on the presence of calcium ions, cellular respiration and free sulphydryl groups. These results suggest that contrary to current belief, all 3 transcobalamins play a role in the transfer of vitamin B₁₂ to tissue cells.

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The Three Transcobalamins in Myeloproliferative Disorders and Acute Leukaemia

Zittoun

Marquet

et al. 1975

Br J Haematol

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The unsaturated vitamin B12 binding capacity of whole serum (UBBC) and of the three transcobalamins (TC) has been studied in patients with various haematological diseases including myeloproliferative disorders (MPD) and acute leukaemia. The binding capacity of TC I and TC III was increased in MPD; TC I being particularly high in chronic granulocytic leukaemia (CGL) and TC III especially raised in polycythaemia rubra vera (PRV) and in infectious leucocytosis. The binding capacity of both TC I and TC III correlated with blood neutrophil count and the ratio TC III/TC I was low in CGL and increased in PRV. TC II was increased in acute myelogenous leukaemia, during remission and blast cell crisis of CGL and in refractory anaemia with excess of myeloblasts but not in acute lymphoblastic leukaemia (ALL). TC II correlated inversely with blood neutrophil count. There is an inverse ratio between TC II and TC I at least in myelogenous leukaemia. These abnormalities are discussed in relation to granulocyte kinetics. TC III and TC I reflect probably the total body granulocyte pool and share some biochemical and immunological properties supporting the view that they have a common origin in the more mature stages of the granulocyte cell line while TC II probably originates partly in more primitive granulocytes.

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Separation of Transcobalamins on Small DEAE-cellulose Columns

Cited by 8 publications

References 1 publication

Comparison of Biochemical Properties of Human Serum B₁₂ Binders Derived from Sephadex and DEAE‐Cellulose Chromatography

Comparison of Biochemical Properties of Human Serum B₁₂ Binders Derived from Sephadex and DEAE‐Cellulose Chromatography

Role of Transcobalamins I, II and III in the Transfer of Vitamin B<sub>12</sub> to Human Bone Marrow Cells <i>in vitro</i>

The Three Transcobalamins in Myeloproliferative Disorders and Acute Leukaemia

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Separation of Transcobalamins on Small DEAE-cellulose Columns

Cited by 8 publications

References 1 publication

Comparison of Biochemical Properties of Human Serum B12 Binders Derived from Sephadex and DEAE‐Cellulose Chromatography

Comparison of Biochemical Properties of Human Serum B12 Binders Derived from Sephadex and DEAE‐Cellulose Chromatography

Role of Transcobalamins I, II and III in the Transfer of Vitamin B<sub>12</sub> to Human Bone Marrow Cells <i>in vitro</i>

The Three Transcobalamins in Myeloproliferative Disorders and Acute Leukaemia

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Comparison of Biochemical Properties of Human Serum B₁₂ Binders Derived from Sephadex and DEAE‐Cellulose Chromatography

Comparison of Biochemical Properties of Human Serum B₁₂ Binders Derived from Sephadex and DEAE‐Cellulose Chromatography