Inhibition of terminal complement complex formation and cell lysis by monoclonal antibodies.

Three monoclonal antibodies (mabs), two against C5 and one against C6, were identified and characterized. They inhibited the generation of the terminal complement complex (TCC) in serum to over 90% as assayed by a sensitive ELISA based on a neoepitope-specific mab, which recognized TCC-integrated C9. The haemolytic function of the TCC was markedly reduced by all three mabs implying that they are directed to epitopes on C5 and C6 which are essential for TCC formation in both the fluid phase and on erythrocyte membranes. Since the generation of C5a was also impaired by these mabs, they may serve as tools in investigations of the sequelae of the generation of C5a and of TCC.

The primary structure and functional properties of the hemoglobins of a ground squirrel (Spermophilus townsendii, Rodentia).

The hemoglobin of the ground squirrel Spermophilus townsendii consists of two components which are present in a ratio of ca. 2:1. The two hemoglobins have identical alpha-chains, but differ in their beta-chains. We present the primary structures of the alpha- and the two beta-globin chains. Following chain separation by chromatography on carboxymethyl-cellulose CM-52, the amino-acid sequences were established by automatic Edman degradation of the globin chains and the tryptic peptides, as well as of a peptide obtained by acid hydrolysis of the Asp-Pro bond of the beta-chains. The two beta-chains differ by only one amino-acid residue, Ala being present in the main and Asp in the minor component in position 58 (E2). The comparison with human hemoglobin showed only 14 exchanges in the alpha-chains but 33 in the beta-chains. Whereas no contact positions are affected in the alpha-chains, we found four such substitutions in the beta-chains, including one heme contact, two alpha 1/beta 1-contacts, and one alpha 1/beta 2-contact. It seems however, that the substitution found in the beta-chains has no effect on the oxygen affinity.

[Pre- and perinatal oxygen transport in mammals: the embryonic hemoglobins of the domestic pig (Sus scrofa domestica)]. / Zur prä- und perinatalen Sauerstoffübertragung der Säuger Die embryonalen Hämoglobine vom Hausschwein (Sus scrofa domestica).

Pigs express two globin genes of beta-type, the epsilon- and chains during their embryonic stage. With the alpha- and epsilon-chains they form four embryonic hemoglobins. We describe here in detail the experimental procedures for sequencing the epsilon- and chains from the pig (Sus scrofa domestica) and we discuss the data with respect to their special functional properties. From the components Gower I and Heide I, we obtained all embryonic chains by chromatography on CM-Cellulose. The sequence of the tryptic peptides of the beta-type chains was established by automatic Edman-Begg degradation. They were aligned by comparison with the corresponding human chains. The epsilon-chains from man and pig differ in 20 positions, porcine and epsilon-chains only in 4 positions. Up to now these have only been found in pigs. A fetal hemoglobin (gamma-chains) was not observed. As a result of this work the sequences of all peptide chains of pig's hemoglobin are determined.

[Primary structure of hemoglobins of the musk rat (Ondatra zibethica, Rodentia)]. / Die Primärstruktur des Hämoglobins vom Bisam (Ondatra zibethica, Rodentia).

The amino-acid sequence for the hemoglobin of the musk-rat (Ondatra zibethica) was determined. The sequence of both chains was established by automatic Edman-Begg degradation of the tryptic peptides and, in the case of the beta-chains additionally of the prolyl-peptide. The complete primary structure of the alpha-chains differs at 22, that of the beta-chains at 36 positions from the adult human hemoglobin. There are no changes in the heme-binding residues, the alpha 1--beta 2 contact positions and the allosteric regulator sites. In the heme-pocket we found beta 44 Ser leads to His and 8 positions changed within the alpha 1--beta 1 contact regions. These changes are discussed with respect to the function of the respective regions.

[Embryonic hemoglobins in mammals: sequences of zeta-, epsilon- and theta chains in domestic pigs (Sus scrofa domestica)]. / Embryonale Hämoglobine der Säuger: Die Sequenzen der zeta-, epsilon- und theta-Ketten vom Hausschwein (Sus scrofa domestica).

The amino-acid sequences of all expressed hemoglobins of the pig embryo are given: Hemoglobin Gower I (zeta 2/epsilon 2), Hemoglobin Gower II (alpha 2/epsilon 2), Hemoglobin Heide I (zeta 2/theta 2) and Hemoglobin Heide II (alpha 2/theta 2). The zeta-, epsilon- and theta-chains were obtained with chromatography on CM-cellulose from isolated hemoglobin components. The primary structure was established by sequencing the tryptic peptides in the sequenator: they were isolated using HPLC. The zeta-chains from pig and human differ in 23, the epsilon-chains in 20 positions. The embryonic globin-gene which express the theta-chains, is a new one in mammals, of epsilon-type and up to now it could only be found in pigs: the amino-acid sequence differ in only 4 positions from the epsilon-chains. Because no gamma-chains (fetal Hb) are expressed the sequences of all hemoglobins (5 hemoglobin chains forming 5 different hemoglobins) of ontogeny in pig are now described.

Hemoglobins, XLI. The embryonic hemoglobins of mammal. A new hemoglobin, hemoglobin Heide II (Hb HeII: alpha 2 epsilon 2), and demonstration of the hemoglobin structure of gower I (zeta 3 epsilon 2) in the pig embryo.

A new hemoglobin is described. The hemoglobins of pig embryonic erythrocytes are separated by DEAE-Cephacel chromatography. In addition to a small amount of adult hemoglobin, four embryonic hemoglobins are obtained. Each of the hemoglobin components is analysed by globin and hemoglobin electrophoresis on polyacrylamide gels. Two of these hemoglobins are previously described as Hb Gower II (alpha 2 epsilon 2) and Heide I (zeta 2 epsilon 2). A tetrameric hemoglobin consisting of alpha and epsilon chains has been detected. Therefore, this new hemoglobin is named, in analogy, as Hb Heide II (alpha 2 epsilon 2). Furthermore, the other hemoglobin tetramer has been unequivocally identified as zeta 2 epsilon 2. This structure corresponds to Gower I which is only suggested in the human embryo.