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K H Scheit

Publications and source records attributed to K H Scheit.

At least 37 records · Page 2Linked to original sources
PubMed1993

Characterization of a full-length cDNA encoding a bovine four tandem-repeat ubiquitin.

Screening of a bovine seminal vesicle cDNA library in lambda gt11 with monospecific anti-ubiquitin IgGs yielded two independent clones pUF4AA and pUD4AA. Sequence analysis of pUF4AA revealed a coding region for a polyubiquitin with four tandem-repeats. We observed a replacement of serine (133) by phenylalanine in the second ubiquitin repeat. The deduced amino acid sequence of pUD4AA, however, possessed the typical serine (133) residue. This serine-phenylalanine replacement could result from allelic polymorphism.

Amino Acid Sequence↗
PubMed1993

The potential use of prostatic secretory protein of 94 amino acid residues (PSP94) as a serum marker for prostatic tumor.

The serum concentrations of prostatic secretory protein of 94 amino acid residues (PSP94) as well as those of prostate-specific antigen (PSA) were determined in 40 patients with established prostatic carcinoma, prior to transurethral resection of the prostate. In a comparison with a control group of healthy men (n = 40) and a group of patients with histologically established benign prostatic hyperplasia (n = 40) no significant differences in PSP94 serum concentrations between the groups were observed. Similarly, correlations of PSP94 serum concentrations with prostatic carcinoma stages or grades were not detected. In contrast, and as expected, PSA behaved as a prostate tumor marker of known sensitivity and specificity. A correlation of PSP94 and PSA concentrations in sera of patients with benign prostatic hyperplasia and/or prostatic carcinoma could not be verified. PSP94 apparently does not fulfill the criteria of a serum marker for monitoring adenomas and/or carcinomas of the prostate.

Adult↗
PubMed1993

Clinical evaluation of a new prostate-specific antigen sandwich ELISA which employs four monoclonal antibodies directed at different epitopes of prostate-specific antigen.

A known radioimmunometric prostate-specific antigen (PSA) test based on monoclonal antibodies, as well as a new PSA-ELISA utilizing 4 monoclonal antibodies directed against different epitopes of PSA were compared in a clinical evaluation. For the investigation, collectives of patient sera from patients with independently diagnosed prostatic carcinoma (PCA) as well as benign prostatic hyperplasia (BPH) were employed. The results of the evaluation demonstrated that although the PSA immunoradiometric test and the PSA-ELISA yielded different numerical values for PSA serum concentrations, they possess comparable diagnostic sensitivities as well as specificities. The nonradioactive PSA-ELISA could therefore substitute the PSA-IRMA in a clinical routine diagnostic of PCA.

Adult↗
PubMed1992

Characterization by cDNA cloning of the mRNA of a highly basic human protein homologous to the yeast ribosomal protein YL41.

From a cDNA library in lambda gt11 derived from poly(A)+ mRNA of human ovarian granulosa cells, a cDNA clone lambda HG12.1, containing an EcoRI insert of 470 bp, was identified. After subcloning of the insert into pUC18, the clone pHG12.1 was obtained and sequenced. The 5'-region of the insert of pHG12.1 was extended by the polymerase chain reaction (PCR) with cloned total cDNA. Assembly of the PCR fragment with the insert of pHG12.1 yielded clone pHG12. From the first open reading frame of pHG12 the amino acid sequence for a polypeptide of 25 amino acid residues (designated HG12) was derived, which was identical in 22 residues with yeast ribosomal protein YL41. It is therefore assumed that HG12 is the first mammalian homolog of yeast ribosomal protein YL41. Transcription of DNA fragments containing the coding region of pHG12 cloned into BluescriptM13, followed by cell-free translation, yielded a polypeptide with an apparent mol.wt. of 14.5 kDa, much larger than the theoretical mol.wt. (3454 Da). The discrepancy between theoretical and apparent mol.wt. was also observed for yeast ribosomal protein YL41. Southern analysis revealed that HG12 is not specified by a single copy gene. Homology for HG12 specific sequences is observed for bovine, porcine and rat species.

Amino Acid Sequence↗
PubMed1992

Characterization by cDNA cloning of the mRNA of a new growth factor from bovine seminal plasma: acidic seminal fluid protein.

A cDNA expression library in lambda gt11 prepared from cDNA derived of seminal vesicle tissue was screened by means of monospecific rabbit anti-aSFP IgG. The sequence of clone pTF21, containing an insert of 668 bp comprised an open reading frame from position 7 to 411 terminated by two stop codons. From this sequence a protein of 134 amino acid residues can be deduced. The mature aSFP was preceded by a signal peptide of 20 amino acids length. The protein sequence contains no signal for N-glycosylation. The molecular weight calculated from the amino acid sequence is 12922 Da. The start codon ATG is part of the sequence AAGATGA which fulfills the criteria of an initiation consensus sequence. The coding region was followed by 257bp of the complete 3'-untranslated region (3'UTR). A putative polyadenylation signal AATAAT, although not of the standard type, is observed at position 650. According to Northern analysis, aSFP mRNA is expressed in seminal vesicle tissue, ampulla and weakly in tissue of epididymis, but not in testis or other bovine tissue. aSFP is specified by a single copy gene. Attempts to detect homologies to known protein sequences were not successful.

Amino Acid Sequence↗
PubMed1992

Construction of a plasmid containing the complete coding region of human elongation factor 2.

A plasmid pUChEF-2 containing the coding sequence as well as the complete 3'-untranslated region (3'UTR) of human EF-2 mRNA was constructed. The plasmid construct was assembled from a cDNA insert of pHGR81 (Rapp et al., (1988) Biol. Chem. Hoppe-Seyler 369, 247-250) comprising the C-terminal portion of the coding region and the 3'UTR, as well as a polymer chain reaction PCR fragment (Rapp et al., (1989) Biol. Chem. Hoppe-Seyler 370, 1071-1075) covering the missing part of the coding region from the amino-terminus.

Base Sequence↗
PubMed1991

Characterization of the gene for the bovine seminal vesicle secretory protein SVSP109.

As part of an attempt to understand androgen-regulation of SVSP109, a bovine seminal vesicle secretory protein of 109 amino acids, we have characterized the bovine SVSP109 gene. The 6.1 kb gene is organized in five exons and four introns. Regulatory sequences involved in regulation of transcription could not be identified by simple sequence homologies. The SVSP109 gene may provide an excellent example for functional properties of exons: exon 1 encodes the entire signal peptide and exon 4 the complete fibronectin type II-domain, responsible for protein-protein interactions.

Amino Acid Sequence↗
PubMed1991

Characterization of a new bioactive protein from bovine seminal fluid.

A new acidic seminal fluid protein (aSFP) was purified from bovine seminal fluid, using anion exchange chromatography and FPLC on MonoQ. The purified aSFP displays a pI of 4.8 and an apparent molecular weight of 14 kDa. Homogeneity of aSFP was demonstrated by FPLC and SDS-polyacrylamide gel electrophoresis. Monospecific anti-aSFP IgGs were employed to characterize aSFP in bovine seminal plasma and seminal vesicle secretion by immuno blot analysis. Proteinchemical characterization of aSFP included amino acid analysis as well as determination of 23 amino acid residues of the N-terminal sequence of aSFP. According to this sequence, aSFP appears to represent a hitherto unknown protein. aSFP stimulated cell division and progesterone secretion of bovine granulosa cells in vitro in a potent and dose dependent manner. aSFP appears to be a potent growth factor with effects on ovarian granulosa cells.

Amino Acid Sequence↗
PubMed1991

Human elongation factor 1 beta: cDNA and derived amino acid sequence.

From a cDNA library in lambda gt11 derived from poly (A+)RNA of human ovarian granulosa cells a cDNA clone lambda HGP34, containing an EcoRI insert of 829 bp, was identified. After subcloning of the insert into pUC18, the clone pHGP34 was obtained and sequenced. The derived amino acid sequence, corresponding to a protein of 225 amino acids, shows a high degree of homology to elongation factor 1 beta (EF-1 beta) of Artemia salina (57%) and known peptide sequences of Xenopus laevis EF-1 beta (86%). We therefore assume that the protein coded for by pHGP34 represents human EF-1 beta. Northern analysis reveals an EF-1 beta specific mRNA of 900 bp. Southern analysis indicates that EF-1 beta in the human genome, like EF-1 alpha, appears to be specified by more than one gene. A high degree of sequence homology for EF-1 beta specific sequences is observed for bovine, rat and mouse species.

Amino Acid Sequence↗
PubMed1991

The evaluation of markers of prostatic function.

The concentrations of three secretory proteins of the human prostate, including prostatic secretory protein of 94 amino acid residues (PSP94), prostate-specific antigen (PSA) and prostatic acid phosphatase (PAP), were measured by enzyme-linked immunosorbent assay (ELISA) in semen from a collective of patients suffering from various inflammatory diseases of the genital tract. In addition, levels of the conventional markers citrate, glucosidase and fructose were determined. As compared with semen from men exhibiting no inflammatory condition, only levels of glucosidase in cases of epididymitis and concentrations of PSP94 in the collective suffering from prostatitis showed significant reductions. The changes in the secretion of PSA, PAP, fructose and citrate in the semen of patients with inflammation of genital tract tissue were not significant at the 95% range of confidence.

Acid Phosphatase↗
PubMed1991

Gene expression and cDNA cloning identified a major basic protein constituent of bovine seminal plasma as bovine monocyte-chemoattractant protein-1 (MCP-1).

P6 is one of the major basic proteins of bovine seminal plasma. Using cell-free translation of poly(A)+RNA from bovine seminal vesicle tissue and monospecific anti-P6-IgGs, we show that P6 is a secretory product of the seminal vesicles. Immunohistochemical experiments supported this finding. Immunoscreening of a lambda gt11 cDNA library derived from seminal vesicle poly(A)+RNA furnished a number of positive cDNA clones, from which clone pH42 was characterized by sequencing. The partial amino acid sequence of a CNBr-fragment of P6 permitted identification of the reading frame of clone pH42 encoding the precursor protein of P6. The P6 precursor contains a signal peptide of 23 amino acids followed by the mature P6 sequence of 76 amino acid residues. The cDNA sequence of pH42 was 80% homologous with that of the human monocyte-chemoattractant protein-1 (hMCP-1). The respective amino acid sequences for the precursor molecules are 72% identical. Northern analysis of seminal vesicle poly(A)+RNA using pH42 as probe probe identified a 0.9-kb P6 mRNA. Stimulation of P6 mRNA expression by phytohemagglutinin in bovine peripheral mononuclear leukocytes suggests that P6 is identical to bovine MCP-1.

Amino Acid Sequence↗
PubMed1990

mRNA of bovine tissue inhibitor of metalloproteinase: sequence and expression in bovine ovarian tissue.

A cDNA library derived from poly(A+)RNA of bovine ovary was screened with a PCR fragment comprising the coding region of human tissue inhibitor of metalloproteinase inhibitor (TIMP). From a number of positive clones, pBGR19, containing a 747 bp insert, was identified and sequenced. The derived amino acid sequence represents that of the precursor of bovine TIMP. Northern analysis reveals a TIMP specific mRNA of 800 bp. Southern analysis indicates that one gene appears to specify bovine TIMP. TIMP mRNA is only weakly expressed in follicular granulosa- and theca cells, whereas luteinization of the follicle is associated with an increase of expression. Expression varies with the stage of the luteal phase; it was highest in stages I and III, but low in stages II and IV of the oestrous cycle.

Amino Acid Sequence↗
PubMed1990

Seminalplasmin, the major basic protein of bull seminal plasma, is a secretory protein of the seminal vesicles.

From the experimental results of three independent methods: (1) indirect immunofluorescence employing monospecific anti-seminalplasmin-IgGs, (2) cell-free translation of poly(A)+ RNA from seminal vesicle and testicular tissue, as well as (3) Northern analysis of poly(A)+ RNA of the latter tissues with a synthetic seminalplasmin-specific antisense DNA probe, it is concluded that the biosynthesis of seminalplasmin occurs in seminal vesicles but not in testis.

Amino Acid Sequence↗
PubMed1990

Characterization of a monoclonal antibody specific for prostatic secretory protein of 94 amino acids (PSP94) and development of a two-site binding enzyme immunoassay for PSP94.

Purified prostatic secretory protein of 94 amino acids (PSP94) was used to generate polyclonal rabbit anti-PSP94 IgG and a murine monoclonal antibody 6B6 (mAB6B6). Both antibodies were highly specific for PSP94 by immunoblotting. Immunohistochemical studies demonstrated the specificity of mAB6B6 for human prostatic epithelial tissue. A two-site binding enzyme immunoassay for the detection of PSP94 was developed using a combination of the antibodies. The sandwich-ELISA yielded satisfactory results when mAB6B6 complexed to peroxidase conjugated goat anti-mouse-IgG (Fc) was incubated simultaneously with the sample. The assay has a dynamic range of 2-30 micrograms/l. This immunoassay was employed to measure PSP94 in male human sera (8 +/- 4 micrograms/l), female human sera (5.7 +/- 3.4 micrograms/l), follicular fluid (3.9 +/- 2.9 micrograms/l) and human seminal plasma (1.02 +/- 0.8 g/l).

Adolescent↗
PubMed1990

Species- and organ-specificity of secretory proteins derived from human prostate and seminal vesicles.

Polyclonal antibodies against semenogelin (SG) isolated from human seminal vesicle secretion and acid phosphatase (PAP), beta-microseminoprotein (beta-MSP), and Prostate-Specific Antigen (PSA) derived from human prostatic fluid, as well as a monoclonal antibody against beta-MSP were used for immunocytochemical detection of the respective antigens in different organs from different species. SG immunoreactivity was detected in the epithelium of the pubertal and adult human and in monkey seminal vesicle, ampulla of the vas deferens, and ejaculatory duct. PAP, beta-MSP, and PSA immunoreactivities were detected in the pubertal and adult human prostate and the cranial and caudal monkey prostate. With the exception of a weak PSA immunoreactivity in the proximal portions of the ejaculatory duct, none of the latter antisera reacted with seminal vesicle, ampullary, and ejaculatory duct epithelium. Among the non-primate species studied (dog, bull, rat, guinea pig) only the canine prostatic epithelium displayed a definite immunoreactivity with the PAP antibody and a moderate reaction with the PSA antibody. No immunoreaction was seen in bull and rat seminal vesicle and canine ampulla of the vas deferens with the SG antibody. The same was true for the (ventral) prostate of rat, bull, and dog for beta-MSP. The epithelium of the rat dorsal prostate showed a slight cross-reactivity with the monoclonal antibody against beta-MSP and one polyclonal antibody against PSA. The findings indicate a rather strict species-dependent expression of human seminal proteins which show some similarities in primates, but only marginal relationship to species with different physiology of seminal fluid.

Acid Phosphatase↗
PubMed1990

Development and evaluation of an enzyme-linked immunoassay for the prostate: specific antigen utilizing two monoclonal antibodies.

PSA is an important tumor-marker for prostatic cancer disease. We developed a sensitive, simple and inexpensive Sandwich ELISA for PSA with two monoclonal antibodies. The precision and reliability of the assay are reflected in the low inter- and intraassay coefficient of variation. PSA was not detectable in sera from normal females (n = 50). Sera from males with different serum levels of PSA (normal males, patients with prostate hypertrophy, prostate cancer patients, n = 79) and 15 prostate cancer patients treated with Zoladex were measured by our ELISA and by a commercially available RIA. The correlation coefficient between these both test systems was close to 1 (r = 0.97).

Antibodies, Monoclonal↗