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K B Pierson

Publications and source records attributed to K B Pierson.

5 recordsLinked to original sources
PubMed1993

Nonisotopic DNA detection system employing elastase and fluorogenic rhodamine substrate.

An alternative fluorescence-based method has been developed for the direct detection of small quantities of DNA in solution. In this system, a serine protease (elastase) is coupled to a DNA oligonucleotide through a disulfide linkage. A bis-(tetraalanine)-derivatized rhodamine molecule BZTAlaR) has been synthesized for use as a substrate. BZTAlaR is nonfluorescent in its derivatized form and shows negligible hydrolysis in solution. Cleavage of the tetraalanyl groups from the rhodamine portion of the molecule restores its fluorescence. Hybridization of the elastase-oligonucleotide conjugate to its target, capture of the conjugate-target complex with streptavidin-coated magnetic beads, addition of substrate, and subsequent detection of the target by fluorescence are accomplished in solution. Hybridization is rapid and specific, with over 90% of a target sequence successfully hybridized and captured. This method exhibits low background and an amplified fluorescent signal over time, resulting in a current detection limit of 0.49 fmol of elastase alone, or 2.64 fmol of conjugate, within 2 h.

Base Sequence↗
PubMed1988

200 Kd neurofilament protein binds Al, Cu and Zn.

200 Kd human and bovine neurofilament proteins were isolated from spinal cord and purified to homogeneity. The purified proteins were shown to be metal-binding proteins which bind at least one mole of Al, one mole of Cu and four moles of Zn. Neither dephosphorylation of the protein nor equilibrium dialysis against 25 mM EDTA, pH 7.0, removed these metals. This is the first report of a human protein which stoichiometrically binds Al.

Aluminum↗
PubMed1985

Isolation and partial characterization of a non-thionein, zinc-binding protein from the liver of rainbow trout (Salmo gairdneri).

A Zn-binding protein (ZBP) was induced in the liver of Donaldson strain rainbow trout by 7 mg/kg i.p. injections of divalent Zn ions. ZBP synthesis was inhibited (74%) by cycloheximide. ZBP had a minimum molecular weight of 16,900 and was composed of two subunits with apparent molecular weights of 9650 and 6050. Purified ZBP contained 4% phenylalanine, 4% isoleucine, 5-6% leucine and 3.48% Zn. Each protein molecule bound nine Zn atoms and contained at least four sulfhydryl groups. Molecular weight, two subunits and the presence of aromatic amino acids suggested that ZBP was not metallothionein.

Amino Acids↗
PubMed1985

Occurrence and synthesis of a non-thionein, zinc-binding protein in the rainbow trout (Salmo gairdneri).

A non-thionein, Zn-binding protein (ZBP) was induced in Donaldson strain rainbow trout (Salmo gairdneri) by 7 mg/kg, i.p. injections of divalent Zn ion. The Sephacryl S-200 used for supernatant fractionation had to be saturated with Zn to recover quantitatively the Zn-ZBP complex. The ZBP was present in liver and kidney, but was absent from gill and spleen. The apparent molecular weights of the liver and kidney ZBP as estimated by gel filtration were 17,300 +/- 1300 (SD; N = 11) and 18,100 (N = 1), respectively. Starvation induced hepatic ZBP synthesis whereas cycloheximide inhibited hepatic ZBP synthesis. The quantity of hepatic ZBP synthesized varied with the temperature of the water in which the trout resided. The maximum quantity of ZBP in the liver following a single 7 mg/kg Zn injection (17 micrograms Zn/g liver wet weight) occurred at 24 hr.

Animals↗