Biomedical subjects

K J Reissner

Publications and source records attributed to K J Reissner.

6 recordsLinked to original sources

PubMed2006

Molecular nodes in memory processing: insights from Aplysia.

Recent research in a variety of systems indicates that memory formation can involve the activation of a wide range of molecular cascades. In assessing this recent work it is clear that no single cascade is uniquely important for all forms of memory, nor is a single form of memory uniquely dependent on a single cascade. Rather, it appears that molecular networks are differentially engaged in the induction of various forms of memory. Despite this highly interactive array of possible cascades, specific 'molecular nodes' have emerged as critical regulatory points in memory formation. Functionally, these nodes can operate in two sequential steps, beginning with a convergence of inputs which coordinately influence the activation state of the node, in which the nature of stimulation determines the dynamics of nodal activity, followed by a divergence of substrate selection, in which the node serves as a gateway that activates specific downstream effectors. Finally, specific nodes can be differentially engaged (i.e. have different 'weights') depending upon the nature and pattern of the activating stimulus. The marine mollusk Aplysia has proven useful for a molecular analysis of memory formation. We will use this system to highlight some of the molecular strategies employed by the nervous system in the formation of memory for sensitization, and we will focus on extracellular signal-related kinase as a candidate node integral to these processes.

Animals↗

PubMed2003

Deamidation and isoaspartate formation in proteins: unwanted alterations or surreptitious signals?

Formation of betalinked Asp-Xaa peptide bonds--isoaspartyl (isoAsp) sites--arise in proteins via succinimide-linked deamidation of asparagine or dehydration of aspartate, reactions which represent a major source of spontaneous protein damage under physiological conditions. Accumulation of atypical isoaspartyl sites is minimized in vivo by the activity of protein L-isoaspartyl O-methyltransferase (PIMT), which regenerates a normal peptide bond. Loss of PIMT has harmful consequences, especially in neurons; thus, formation of isoAsp sites and their subsequent correction by PIMT is widely believed to constitute an important pathway of protein damage and repair. Recent evidence is mounting, however, that deamidation and isoaspartate formation may, in some instances, constitute a novel mechanism for intentional modification of protein structure. Herein we describe the mechanism of Asx rearrangement, summarize the evidence that PIMT serves an important repair function, and then focus on emerging evidence that deamidation and isoAsp formation may sometimes have a useful function.

Amides↗

PubMed1998

Isoaspartate in chrondroitin sulfate proteoglycans of mammalian brain.

Mammalian brain contains a high mass protein (HMAP) that is unusually rich in atypical L-isoaspartyl (isoAsp) linkages. HMAP has now been purified from bovine brain by anion exchange, hydroxylapatite, and size exclusion chromatography. It is self-aggregating, acidic, and soluble in 5% trichloroacetic acid. Treatment with chondroitinase ABC eliminates the self-aggregation of HMAP and generates several distinct core proteins with estimated masses of 350-450 (doublet), 180, and 100 kDa, indicating that it is composed mainly of chondroitin sulfate proteoglycans (CSPGs). Most of the isoAsp resides in the 350-450-kDa core protein, which was identified by immunoblotting as phosphacan, a CSPG abundant in adult brain. The regional distribution and developmental profile of HMAP in rat brain support this identification. The 180-kDa core protein contains a tenascin-R-related molecule, consistent with recent observations that phosphacan forms a tight complex with tenascin-R. The average phosphacan molecule in adult brain contains at least seven isoAsp sites. Molecular heterogeneity due to isoAsp may explain some of the complex binding properties phosphacan exhibits with its natural ligands. Formation of isoAsp may be important in the roles that phosphacan and other CSPGs play in development of the nervous system.

Animals↗

PubMed1998

Regulation of platelet heparanase during inflammation: role of pH and proteinases.

Heparan sulfate is rapidly degraded by an endoglycosidase (heparanase) secreted by activated platelets. Since the cleavage and release of heparan sulfate would profoundly alter the local physiology of the endothelium, platelet heparanase activity should be tightly regulated. Consistent with this hypothesis, platelet heparanase was found to degrade endothelial cell heparan sulfate at pH 6.0 but not at pH 7.4, even though 25% of maximum activity was detected at pH 7.4. Loss of heparanase activity occurred rapidly (t1/2 is approximately equal to 20 min) and reversibly at physiologic pH but did not occur at acidic pH (<7.0). Inactivation of heparanase at pH 7.4 did not affect heparin binding and was reversed by 0.5 M NaCl or by heparan sulfate but not by chondroitin sulfate, suggesting inactive heparanase could be tethered on cell surfaces and the function regulated by heparan sulfate. Heparanase was gradually inactivated by trypsin and urokinase (t1/2 = 5 h) but resisted cleavage by leukocyte cathepsin G, leukocyte elastase, plasmin, and thrombin. These findings are consistent with a model in which platelet heparanase is active at the low pH of inflammation but inactive under physiologic conditions preventing inadvertent cleavage of heparan sulfate and loss of physiologic functions of endothelial cells.

Animals↗

PubMed1998

Genotypic heterogeneity and phenotypic variation among patients with type 2 Gaucher's disease.

Gaucher's disease, the inherited deficiency of glucocerebrosidase, manifests with vast phenotypic variation. Even among patients with type 2 (acute neuronopathic) Gaucher's disease, there is a spectrum of clinical presentations. DNA samples from 14 patients with type 2 Gaucher's disease with a course ranging from intrauterine death at 22 wk of gestation to survival until age 30 mo were studied. L444P was the only common mutation identified, found in 15 patients' alleles. Sequencing of genomic DNA amplified by long template PCR revealed that mutation L444P occurred as a single point mutation in seven mutant alleles and as part of a recombinant allele in eight mutant alleles. Two patients had a deletion of 55 bp in exon 9; in one patient the deletion was part of a recombinant allele, and in a second the deletion occurred alone. Direct sequencing identified R120W on one allele, P415R on another, and one fetus was homoallelic for a deletion of a C nucleotide at codon 139 in exon 5. Eight of the mutant alleles remain unidentified. Northern blots revealed an appropriately sized mRNA in all except one of the patients studied. Of the 14 type 2 Gaucher patients, three had hydrops fetalis and died in utero or at birth, five had congenital ichthyosis, and seven survived 5 mo or more. Patients who died in the neonatal period had decreased protein detected by Western blot, regardless of genotype observed. These studies demonstrate that genotypic heterogeneity exists in patients with type 2 Gaucher's disease, even among infants with the most severe phenotypes.

Abortion, Spontaneous↗

PubMed1998

Fate of antigen in xenotransplantation: implications for acute vascular rejection and accommodation.

Antigen down-modulation plays a critical role in xenotransplants involving humoral responses against the Forssman antigen and may play a role in the long-term survival of ABO-incompatible allografts. The present study investigates the fate of porcine antigens in pig-to-primate xenotransplantation. Human antibodies bound to the glycocalyx of cultured porcine aortic endothelial cells as judged by electron microscopy and were shed from the cell surface in a complex with fibronectin, a glycoprotein that is found in the apical membrane glycocalyx of cultured cells. Antibody was shed in a metabolically dependent process with a t(1/2) of 2 to 3 hours. However, the amount of antigen on the cell surface did not change appreciably within 24 hours, suggesting that antigen modulation did not occur. Over the ensuing days, antigen expression decreased, although the change was always less than 50% of baseline. Changes in antigen expression were due for the most part to changes in expression of alpha-galactosyl residues. Consistent with results obtained in vitro, antigen expression in porcine organ transplants remained at approximately the baseline level as determined by immunofluorescence analysis of IgM binding to graft endothelium. If, as these results suggest, antigen is not down-modulated in pig-to-primate xenotransplantation, then therapies aimed at prolonged xenograft survival must focus on antibody or genetic manipulation of antigen expression.

Animals↗