Hemoglobin-based blood substitutes: characterization of five pyridoxal 5'-phosphate derivatives of hemoglobin

Journal of Chromatography
M J McGarrityJ C Hsia

Abstract

Six fractions (I-VI) of pyridoxal 5'-phosphate (PLP) hemoglobin (Hb), prepared by the method of De Venuto and Zegna [J. Surg. Res., 34 (1983) 205] have been isolated and purified by anion-exchange high-performance liquid chromatography (HPLC). Total phosphate analyses indicate that I is HbA, II and III are double-labelled, IV and V are tetra-labelled and VI contains 6 mol of phosphorus per mol of hemoglobin. The purified components have been resolved into their alpha and beta chains by preparative reversed-phase HPLC using a macroporous C4 support. Phosphate analyses indicate that the beta chains of II, III and IV each contain one phosphate per chain while the beta chains of V and VI each contain two phosphates. The alpha chains of IV and VI were found to be monophosphate-labelled. Reversed phase HPLC analysis of the tryptic peptides of the beta chains indicates that the label is bound exclusively to the 1-valine residue in II, III and IV while both the 1-valine and the 82-lysine are labelled in V and VI. Similarly, modification of the 1-valine residue of the alpha chains of IV and VI was detected. Components II and III have the same molecular formula. Evidence is presented which shows that they are interconvertible and that th...Continue Reading

References

Jan 10, 1986·Journal of Chromatography·J C Hsia, S S Er
Jul 1, 1973·Analytical Biochemistry·R C Williams, K Y Tsay
Jan 1, 1981·Methods in Enzymology·E Bucci
Mar 1, 1983·The Journal of Surgical Research·F DeVenuto, A Zegna
Jul 1, 1983·International Journal of Peptide and Protein Research·A S AcharyaJ M Manning
Feb 1, 1981·The Journal of Surgical Research·L R SehgalG S Moss

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Citations

Mar 1, 1990·Biulleten' eksperimental'noĭ biologii i meditsiny·E P ViazovaA I Zeĭnalov
Jan 1, 1988·Biomaterials, Artificial Cells, and Artificial Organs·J C Hsia, S S Er
May 9, 2000·Journal of Investigative Surgery : the Official Journal of the Academy of Surgical Research·H I FriedmanR Bynoe

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