Purification of Murine and Feline Type-C Virus
Envelope Polypeptides as Micellar Protein
Josef Schneider*, Heinz Falk **,
and Gerhard Hunsmann*
Z. Naturforsch. 36 c, 353-356 (1981);
received January 13, 1981
Mammalian Type-C Viruses, Envelope Polypeptides, Puri
fication, Micellar Complexes, PeptideMaps
A technique originally described for the isolation of
Friend leukaemia virus envelope polypeptides  yields
equivalent structures from Moloney leukaemia, AKR and
BALB/c xenotropic virus as well as feline leukaemia virus.
of the core polypeptides pl8 and p24 of HIV-2, three HIV-1 and five SIV isolates. Each peptidemap was distinguishable, and differences are most prominent between the HIV-1 group and the
SIVmac/SIVsm group. HIV-2 is very similar to SIVmac and SIVsm. The three SIVagm isolates
form a more heterogeneous group. The p24s of all SIVagms are more similar to the p24s of
HIV-1, but with respect to pl8, one isolate is similar to HIV-1, while the two others are more
related to SIVmac, SIVsm, and HIV-2.
The human immunodeficiency virus (HIV) is the
trometer (Thermo Electron, Germany). Mascot
software (Matrixscience) was used for screening the
database NCBInr for proteins corresponding to the
peptidemaps that were obtained.
Results: EBCs from 17 young healthy non-smoking
donors were collected. Different methods for concen-
trating protein were compared in order to optimize
EBC preparations for proteomic analysis. The proce-
dure that was chosen allowed identification of pro-
teins exhaled by healthy people. The major proteins
in the condensates were cytoskeletal keratins.
Another 12 proteins were identified in EBC
The effect of C-terminal amidation on the antimicrobial and hemolytic activities of antimicrobial peptides was studied using three cationic peptides which form amphiphilic α-helices when bound to membranes. The natural antimicrobial peptide PGLa, the designer-made antibiotic MSI-103, and the cell-penetrating "model amphipathic peptide" (MAP) are all amidated in their original forms, and their biological activities were compared with the same sequences carrying a free C-terminus. It was found that, in general, a free COOH-terminus reduces both the antimicrobial activity and the hemolytic side effects of the peptides. The only exception was observed for MSI-103, whose antimicrobial activity was not decreased in the acid form. Having shown that the therapeutic index (TI) of this novel peptide is significantly higher than for the other tested peptides, with high antibiotic activity and little undesired effects, we suggest that it could be a useful starting point for further development of new peptide antibiotics.
Filarial thioredoxin and transglutaminase are enzymes that are secreted throughout the lifecycle of the parasites which are mandatory for the survival of the parasite. They are reported to be promising vaccine candidates, yet the limitation factors of these proteins to be developed as vaccines is their homology they share with the host proteins. Hence immunodominant epitopes from these proteins were constructed as peptides and immunised in mice model with Muramyl dipeptide (MDP) as adjuvant. Immunodominant epitopic portions from Filarial thioredoxin and transglutaminase which are non-homologous with host proteins were constructed as Multi Antigen Peptide (MAP) and assembled in an inert lysine core. The synthesised MAP was immunised with MDP as adjuvant in Balb/c mice model, humoral and cellular immune response were studied. Antibody titre levels for TT MAP with MDP was in par with alum as adjuvant in mice models. T cell responses of TT MAP with MDP showed a balanced TH1/TH2 response. The TH1 cytokines namely IL-2 and IFN-ɤ were also higher in TT MAP immunised groups with MDP as adjuvant whereas alum immunised groups was TH2 biased. TT MAP admixed with MDP as adjuvant proves to be safe in mice model. Further vaccination studies are underway in permissive animal models to determine the role of TT MAP with MDP as adjuvant in protective immunity against W. bancrofti and B. malayi infections.
the absence of
Dialysis of native and inhibited trypsin: Solu
tions of native and inhibited trypsin were dialysed
over a period of 60 hours. Separate samples were
taken at suitable intervals of time and assayed for
enzyme activity in the usual manner.
Preparation of peptidemaps (finger prints) of
native and inhibited trypsin: A sample of lyo-
philized inhibited enzyme (6 —8 mg) from which
excess dibenamine had been removed by diafiltra-
tion was dissolved in a solution of ammonium bi
carbonate (0.5ml; 0.1m; pH 8.7). This solution
sex specificity are not known for cer-
tain. Using triacetyloleandomycin-metabolite com-
plex formation, testosterone hydroxylase assays and
immunoblots from peptidemaps, we obtained results
suggesting that in liver microsomes from mature rats,
at least three, possibly four CYP3A proteins are ex-
pressed: one is present in females, another is male-
specific, and one or two additional CYP3A proteins
are inducible by phenobarbital, steroids, and tri-
Key terms: P450 3A multiplicity, sex specificity, peptidemap.
The P450 cytochromes are members
properties of the two proteases. Such comparative
investigations are also of considerable interest for
classification and phylogenetic aspects within the
The following experiments were performed: amino
acid analysis, polyacrylamide gel electrophoresis,
immunodiffusion, determination of pH-optima and
TV-terminal amino acids, investigation of action
towards selected polypeptides and proteins as
substrates and preparation of peptidemaps of
tryptic and peptic digests of the two proteases.
Materials and Methods
The two enzymes were
. Res. 8: 455–466.  Markovič, O. & Patočka, J. 1977. Action of iodine on the tomato pectinesterase. Experientia 33: 711–712. http://dx.doi.org/10.1007/BF01944143  Markovič, O. & Rexová, L’. 1957. Bestimmung von Scopolamin neben einem Überschuss von Morphin und Äthylmophin. Chem. Zvesti 11: 192–197.  Markovič, O. & Rexová, L’. 1963. Untersuchung von Komponenten verschiedener Bienengiftarten. Chem. Zvesti17: 676–684.  Markovič, O. & Sajgó, M. 1977. Terminal amino acids, peptidemap and amino acid composition of one form of tomato pectinesterase. Acta
be included in the zygote or the shell layers.
Fig. 3 Peptidemaps of albumen gland in E. peliomphala
during feeding period. Experimental"conditions are
described in the text.
5) Peptidemap of albumen gland and egg
The peptidemaps of the albumen gland were observed by
the method of O'Farrell using Coomassie brilliant blue
R-2 50. The specific peptidemap of the albumen gland in
E. peliomphala was shown in Fig.3. The peptidemaps of
the albumen gland were expressed essentially according
to the principle described for that of the egg proteins