The hydrophobic character of the membrane-bound enzyme, adenylate cyclase, was investigated by examining its interaction with a variety of noncharged hydrophobic resins consisting of long chain alkyl groups linked to Sepharose via ether bonds. Either dodecyl or cetyl resins effected the separation of cyclase from more hydrophobic proteins while octyl-, hexyl-, and butyl-Sepharose did not preven...

A method was developed to separate and quantitatively determine two different triglyceride lipase activities in human postheparin plasma: hepatic triglyceride lipase (H-TGL) and lipoprotein lipase (LPL). Affinity chromatography on heparin-Sepharose columns was used for the separation. Rechromatography of purified H-TGL on heparin-Sepharose resulted in recoveries of 74 and 97% of these enzyme ac...

The paper reports an efficient procedure for the extraction and purification of R-phycoerythrin (R-PE) from marine red algae Amphiroa anceps. A. anceps was extracted in 50 mM phosphate buffered saline and precipitated by (NH4)2SO4. The R-PE was isolated by ion-exchange chromatography loaded with Q-Sepharose Fast Flow, which was developed by linear ionic strength gradients. Then it was purified ...

Line 10 guinea pig carcinoma cells cultured in serum-free medium for 4 hr elaborate plasminogen activator (PA) activity that remained in the supernatant after ultracentrifugation (100,000 X g, 90 min). PA activity in line 10 conditioned medium occurred in both active and cryptic forms. The vast majority of active PA adsorbed to lysine-Sepharose and could be eluted at low pH as several activitie...

When rat cerebral-cortex membranes were labelled with [3H]propylbenzilylcholine mustard ([3H]PrBCM), a single protein of Mr 68,000 was found to carry the atropine-sensitive covalent label. After trypsinolysis of the receptors solubilized in 0.075% SDS, the resulting fragments were submitted to size analysis in combination with wheat-germ agglutinin (WGA)-Sepharose and organomercurial-agarose ch...

The binding of tissue-type plasminogen activator (t-PA) to fibrin is mediated both by its finger domain and by its kringle-2 domain. In this report, we investigate the relative affinities of these domains for lysine. Human recombinant t-PA deletion-mutant proteins were prepared and their ability to bind to lysine-Sepharose was investigated. Mutants containing the kringle-2 domain bound to lysin...

Pyran covalently linked to cyanogen bromide-activated Sepharose has been shown to be an effective affinity matrix for several viral DNA polymerases. Differential salt elution of viral compared with cellular polymerases, as well as substrate elution, suggests the affinity nature for the matrix. Unlike some other affinity systems described, pyran-Sepharose is totally resistant to nuclease digesti...

A cyanide biosensor based on a pH-sensitive p-doped electrolyte-insulatorsemiconductor (EIS) structure with an immobilised enzyme (cyanidase) is realised at the laboratory scale. The immobilisation of the cyanidase is performed in two distinct steps: first, the covalent coupling of cyanidase to an N-hydroxysuccinimide(NHS) activated Sepharose gel and then, the physical entrapment of NHS-activat...