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Y C Pan

Publications and source records attributed to Y C Pan.

108 records · Page 6Linked to original sources
PubMed1982

Structure of dihydrofolate reductase: primary sequence of the bovine liver enzyme.

The primary structure of dihydrofolate reductase from bovine liver has been established by Edman degradation of the intact carboxymethylated protein and of peptides obtained from the protein by the action of cyanogen bromide, trypsin, and the protease from Staphylococcus aureus, respectively. Since separation of some of the peptide mixtures by classical methods proved impossible, new systems were developed for the use of high-performance liquid chromatography to separate such mixtures. Some of the cleavage procedures used to obtain peptides gave atypical results at certain peptide bonds. The results are discussed in terms of the residues involved in these unexpectedly resistant or sensitive bonds. The sequence of the bovine liver enzyme is compared with those published for the enzyme from other sources, and known or probable functions of invariant residues are described. Sequences of vertebrate and bacterial reductases are compared and contrasted, and a possible role is considered for the residues which are invariant in bacterial reductases, but different in vertebrate reductases, in determining the selective inhibitory action of trimethoprim on bacterial reductases.

Amino Acid Sequence↗
PubMed1982

Structural analysis of adult and larval isozymes of sn-glycerol-3-phosphate dehydrogenase of Drosophila melanogaster.

Compositional analysis of the soluble tryptic peptides representing about 70% of the 293 residues of sn-glycerol-3-phosphate dehydrogenase in Drosophila melanogaster reveals a single peptide difference between the sn-glycerol-3-phosphate dehydrogenase adult (GPDHF-1) and larval (GPDHF-3) isozymes. This peptide was shown to be the carboxyl terminus by sequence determination and by carboxypeptidase A digestion of the native protein. For GPDHF-1, the sequence of the COOH-terminal tryptic peptide is Asn-His-Pro-Glu-His-Met-Gln-Asn-Leu-COOH, while that of GPDHF-3 is Asn-His-Pro-Glu-His-Met-COOH.

Amino Acid Sequence↗
PubMed1982

Structure of secretory protein IV from rat seminal vesicles.

The complete amino acid sequence of rat SVS-IV protein, consisting of 90 residues, has been determined. The sequence of rat SVS-IV protein is the first seminal vesicle secretory protein determined and it does not show any homology with other known protein sequences. The secondary structure of SVS-IV protein is analyzed by methods of Fasman and Chou indicates that this protein contains 53% alpha-helix, 36% beta-turn and 11% random coil.

Amino Acid Sequence↗
PubMed1980

Amino acid sequence studies on T4 gene 32 DNA-binding proteins.

The amino acid sequence of a total of 281 residues (85%) out of 333 residues from T4 gene 32 protein has been determined. The unusual characteristics of the amino-terminal A and carboxyl-terminal B regions are discussed in relation to their possible roles in DNA replication and genetic recombination.

Amino Acid Sequence↗
PubMedDate not supplied

Purification and sequencing of a rat intestinal 22 amino acid C-terminal CCK fragment.

Fractionation on Sephadex G50 gel of methanol extracts of rat intestine revealed two molecular forms of cholecystokinin (CCK) of about equal immunopotency: one form has an elution volume between CCK33 and CCK12; the other elutes in the salt region as does authentic CCK8. Purification and sequencing have demonstrated that the smaller molecular form is CCK8 with a sequence identical to the pork and sheep CCK8's that had previously been sequenced. Purification and sequencing of the larger molecular form reveals that it is a 22 amino acid C-terminal CCK fragment identical with pig CCK22 except that glycine instead of serine is present at the nineteenth residue from the C-terminus. This sequence is consistent with that predicted by cloned cDNA encoding preprocholecystokinin from a rat medullary thyroid carcinoma. CCK22 has not previously been reported to be a prominent molecular form in either pig or dog intestines.

Amino Acid Sequence↗
PubMedDate not supplied

Sequences of gastrins purified from a single antrum of dog and of goat.

Heptadecapeptide gastrins (G17) have been purified and sequenced from a variety of species. However, progastrin (G34) sequences have been determined only for pig and human from purified peptides and for rat from cDNA. Since G34 in most species accounts for only approximately 5% of total antral gastrin, micropurification techniques must be employed to avoid the need for large quantities of antral tissue. Efficient purification methodology yielded 1.5 and 1.3 nmol of G34 from the antrum of a single goat and of a single dog, respectively. The N-terminal pyroglutamyl residues were enzymatically removed and the peptides were sequenced through to the proximity of their COOH-termini. The COOH-terminal sequences of goat and dog G34 were confirmed by sequencing the corresponding deblocked G17 from each animal. The previously published dog G17 sequence was shown to be incorrect. The sequences for dog and goat G34 are: Dog less than ELGLQGPPQLVADLSKKQGPWMEEEEAAYGWMDF# Goat less than ELGLQDPPHMVADLSKKQGPWVEEEEAAYGWMDF# Dog and goat gastrins differ in 3 sites in the 17 amino acid NH2-terminus and only a single site in G17 (the sites of differences are underlined). The ratio for sulfated to non-sulfated antral G17 is 9:1 for the goat and 1:9 for the dog.

Amino Acid Sequence↗
PubMedDate not supplied

Guinea pig pancreatic polypeptide: structure and pancreatic content.

Guinea pig (GP) pancreatic polypeptide (PP) has been purified from an acid-alcohol extract of 6 GP pancreata by a series of 3 HPLC steps. The sequence for GP PP as compared with that of beef and human is shown: (Sequence: see text). In a single GP pancreas weighing 2.4 g the total PP content was 1.0 nmol and the total glucagon content 61 nmol; in a single dog pancreas weighing 35 g the total PP was 385 nmol and the total glucagon 81 nmol. The relatively low content of PP in GP pancreas is consistent with the fact that the GP lacks a ventral pancreas, the region in which PP is found in highest concentration. The high glucagon content of GP pancreas is consistent with that reported in earlier studies.

Amino Acid Sequence↗
PubMedDate not supplied

Gene expression and apoptosis in the spinal cord neurons after sciatic nerve injury.

To demonstrate a dependence of spinal cord motoneurons on the communication with their targets, the expression of immediate early gene c-fos and neurotrophin genes in the lumbar (L3-L6) spinal cord neurons was examined in Sprague-Dawley rats (male > or = 9-weeks-old) with unilateral sciatic nerve transection. Using in situ hybridization, we detected the expression of c-fos mRNA in the motoneurons of the spinal cord segments within 45 min to 3 h of peripheral nerve transection (n = 4 in each time point). The expression of c-fos mRNA was also correlated positively with the expression of Fos antigen using immunohistochemistry, while no change in calbindin and parvalbumin antigens were noted. The expression of BDNF mRNA increased at 90 min after sciatic nerve transection. However, no detectable enhancement in the expression of NGF mRNA was observed. DNA fragmentation in neurons was observed using the incorporation of digoxigenin-dUTP by terminal transferase into 3'-OH terminals of DNA fragments in the ipsilateral section of the spinal cords 48h after nerve injury. Nuclei that exhibited DNA fragmentation were not observed in the spinal cord of the control animals. Lastly, we observed that the majority of astrocytes did not have DNA fragmentation. Because the detection of DNA fragmentation using this assay is one of early detections of apoptosis or programmed cell death, the result suggested we could detect early cell death in spinal cord, and indicated a target dependence of the neurons in the spinal cord after transection of sciatic nerve.

Animals↗
PubMedDate not supplied

Cloning and expression of murine interleukin-1 cDNA in Escherichia coli.

Interleukin-1 (IL-1), a peptide hormone produced by activated macrophages, possesses the ability to modulate the proliferation, maturation and functional activation of a broad spectrum of cell types and may play a major role in the initiation and amplification of immune and inflammatory responses through its action on these diverse cell populations. IL-1 exhibits microheterogeneity in terms of its relative molecular mass (Mr, 13,000-19,000) and charge properties, and although murine IL-1 has been purified and some of its basic structure-function relationships have been elucidated, it has proved difficult to prepare sufficient amounts of IL-1 for direct and detailed sequence and structural studies. Here we report the cloning, sequence analysis and expression of murine IL-1 cDNA in Escherichia coli. The IL-1 cDNA codes for a polypeptide precursor of 270 amino acids. Biologically active IL-1 was produced in E. coli by expressing the carboxy-terminal 156 amino acids of the IL-1 precursor.

Amino Acid Sequence↗
PubMedDate not supplied

Identity of tumour necrosis factor and the macrophage-secreted factor cachectin.

In mammals, several well-defined metabolic changes occur during infection, many of which are attributable to products of the reticuloendothelial system. Among these changes, a hypertriglyceridaemic state is frequently evident, resulting from defective triglyceride clearance, caused by systemic suppression of the enzyme lipoprotein lipase (LPL). We have found previously that macrophages secrete the hormone cachectin, which specifically suppresses LPL activity in cultured adipocytes (3T3-L1 cells). When originally purified from RAW 264.7 (mouse macrophage) cells, cachectin was shown to have a pI of 4.7, a subunit size of relative molecular mass (Mr) 17,000 and to form non-covalent multimers. A receptor for cachectin was identified on non-tumorigenic cultured cells and on normal mouse liver membranes. A new high-yield purification technique has enabled us to determine further details of the structure of mouse cachectin. We now report that a high degree of homology exists between the N-terminal sequence of mouse cachectin and the N-terminal sequence recently determined for human tumour necrosis factor (TNF). Purified cachectin also possesses potent TNF activity in vitro. These findings suggest that the 'cachectin' and 'TNF' activities of murine macrophage conditioned medium are attributable to a single protein, which modulates the metabolic activities of normal as well as neoplastic cells through interaction with specific high-affinity receptors.

Adipose Tissue↗
PubMedDate not supplied

Elevated expression of the neutrophil calcium-binding protein, MRP-14, in metastasis-enhancing neutrophils.

Tumor-elicited neutrophils (tcPMN) purified from 13762NF mammary adenocarcinoma tumor-bearing rats enhanced metastasis of syngeneic cells when co-injected intravenously; whereas, circulating (cPMN) and phorbol esteractivated (PMA-PMN) neutrophils did not [Welch et al. (1989) Proc. Natl. Acad. Sci. 86:5859-63]. We hypothesized that differential protein expression was responsible for functional differences between the neutrophil subtypes. Two-dimensional polyacrylamide gel electrophoresis was used to compare neutrophils (cPMN, PMA-PMN) purified from the peripheral blood of healthy, syngeneic nontumor-bearing rats, to tcPMN collected from rats with highly metastatic [clone MTLn3, subclone MTLn3(T44).5] or poorly metastatic [subclone MTLn3(T44).11] tumors growing in the mammary fat pads. Quantitative differences in polypeptide expression were observed between these functionally distinct PMN populations. Compared to cPMN, expression of a M(r) approximately 38.8 kDa (pl approximately 8) polypeptide was similar in tcPMN collected from poorly metastatic tumor-bearing rats, higher in PMA-PMN, and further increased in tcPMN from rats with highly metastatic tumors. Expression of two polypeptides, M(r) approximately 14.1 kDa (pl approximately 6) and M(r) approximately 43.3 kDa (pl approximately 5), was greater in tcPMN from rats with highly metastatic tumors compared to cPMN, PMA-PMN, or tcPMN from rats bearing poorly metastatic tumors. The latter two polypeptides thus appeared to be specifically increased in tcPMN from rats bearing highly metastatic tumors. Because it was most abundant and displayed the greatest differences between PMN subtypes, the M(r) approximately 14.1 kDa protein was further analyzed. Tryptic digests followed by internal sequence analyses of resulting peptide fragments revealed that the M(r) approximately 14.1 kDa contained amino acid sequences that were identical to those of MRP-14, a 14 kDa neutrophil calcium-binding protein belonging to the S-100 protein family of calcium-binding proteins. These results suggest a novel function for MRP-14 and suggest that MRP-14 may represent a marker for distinguishing phenotypically distinct subpopulations of neutrophils, particularly tcPMN with metastasis-enhancing abilities.

Amino Acid Sequence↗