Hydrolysis of succinyl-trialanine p-nitroanilide by two enzymes associated with human high-density lipoproteins

Succinyl-trialanine p-nitroanilide (Suc-Ala3-pNA), a synthetic substrate for the determination of elastase activity, was hydrolyzed in sequence by two enzymes that were found to be associated with human high-density lipoproteins. The enzymes involved in the sequence of reaction were separated by ion-exchange chromatography from apo-lipoprotein A-I and A-II, major apoproteins of high-density lipoproteins. One, designated as fraction MK, cleaves Suc-Ala3-pNA to succinyl-dialanine and alanine p-nitroanilide (Ala-pNA), and the other, designated as fraction U, cleaves Ala-pNA to alanine and p-nitroaniline. Fraction MK was inhibited by dithiothreitol, EDTA, and 1,10-phenanthroline, whereas fraction U was inhibited by 1,10-phenanthroline and bestatin. In addition to these findings, fraction MK also hydrolyzed 2,4-dinitrophenyl-prolyl-glutaminyl-glycyl-isoleucyl-alanyl-glycyl-glutaminyl- arginine (DNP-octapeptide), a specific substrate for the determination of vertebrate collagenase. Neither native elastin nor native collagen was hydrolyzed by a mixture of the two enzymes. Fraction U was very similar to aminopeptidase M with respect to its enzyme characteristics studied.     

Polymorphisms of apolipoprotein C-III in two cases with sialidase deficiency

Since sialidase is thought to be one of the enzymes which are concerned with the polymorphic forms of apolipoprotein C-III depending on the contents of terminal sialic acid, the polymorphic forms of apolipoprotein C-III of very low density lipoprotein and apolipoprotein C-III levels were studied in two cases of sialidosis due to sialidase deficiency with or without β-galactosidase deficiency. The diagnosis was established by the defect of those enzymes in the leucocytes and cultured fibroblasts. But, no significant differences in polymorphic form of apolipoprotein C-III were observed between these two patients and controls.     

Flower-Inducing Activity of Water Extracts of Various Plant Species, in Particular Pharbitis nil

The crude water extracts of leaves of many plant species belongingto Spermatophyta and some belonging to Bryophyta induced floweringof Lemna paucicostata 151 (PI51) under continuous light, atthe concentrations equivalent to 0.1 to 10 mg fr wt leaf per10 ml culture medium (mg fr wt/10 ml). The extract of Salvinia(Pterydophyta) added together with the extract of Lemna at aconcentration lower than that necessary to cause flowering alsoinduced flowering. The activity of the water extracts of someplants varied considerably from experiment to experiment dueto unknown factors, but the extracts of Pharbitis nil strainViolet, a sensitive short-day plant, always showed a high activity,as did the extracts of Lemna paucicostata reported previously. The extract of Pharbitis cotyledons induced flowering of P151even at 0.3 mg fr wt/10 ml, and significantly promoted floweringof L. paucicostata 441 and 6746 at 1–3 mg fr wt/10 ml.Ex-udate from the cuttings of the seedlings was also active.However, neither the activity of the water extract nor thatof the exudate could be correlated with photoperiodic floralinduction. On the other hand, the extract of leaves or cotyledonshad higher activity (on a fr wt basis) than that of other organs,and the activity of the extract of cotyledons changed with theirage roughly in parallel with their photoperiodic sensitivity. (Received April 17, 1989; Accepted August 10, 1989)     

Physical Basis of Flower-opening in Pharbitis nil

Flower buds of Pharbitis nil cut from plants growing in thefield open rapidly when subjected to darkness (20–25°C)or low temperature (20°C) in light. Petals of the buds arethe sites of photo- and thermo-perception; flower-opening iscaused mainly by the epinasty of petal midribs. ¹Dedicated to Professor Dr. Erwin Bunning on the occasion ofhis 75th birthday. (Received October 23, 1980; Accepted December 15, 1980)     

Sequence Analysis of the groESL-cotA Region of the Bacillus subtilis Genome, Containing the Restriction/Modification System Genes

We have determined a 35-kb sequence of the groESL-gutR-cotA(45°–52°) region of the Bacillus subtilis genome.In addition to the groESL, gutRB and cotA genes reported previously,we have newly identified 24 ORFs including gutA and fruC genes,encoding glucitol permease and fructokinase, respectively. Theinherent restriction/modification system genes, hsdMR and hsdMM,were mapped between groESL and gutRB, and we have identifiedtwo open reading frames (ORFs) encoding 5-methylcytosine formingDNA methyl transferase and an operon probably encoding a restrictionenzyme complex. The unusual genome structure of few ORFs andlower GC content around the restriction/modification genes stronglysuggests that the region originated from a bacteriophage integratedduring evolution.     

Difficulties of molecular dissociation of Clostridium botulinum type G progenitor toxin

Clostridium botulinum type G progenitor toxin was chromatographed on DEAE-Sephadex and Q-Sepharose equilibrated with 0.05 M Tris-HCl buffer, pH 8.0, containing 0.2 M urea. The toxin was eluted in a single protein peak from DEAE-Sephadex, but it was eluted in four protein peaks from Q-Sepharose; the third peak was toxic and the others were nontoxic. The third peak, appearing to be the toxic component, had a molecular mass of 150,000. In SDS-polyacrylamide gel electrophoresis, purified type G progenitor toxin migrated in six bands, with molecular masses of 150,000, 140,000, 58,000, 10,800, 10,600, and 10,400. Type G progenitor toxin may be composed of a toxin component with a molecular mass of 150,000 and a nontoxic component in a manner similar to progenitor toxins of other types. Type G toxic component, whether it was reduced or not, migrated in a single band to the same relative positions in SDS-PAGE; type A toxic component reduced with 2-mercaptoethanol migrated in two bands.