Algal virus lysin - summary (extensive summary)

Algal virus lysin - summary

Basics of measuring lysin activity - lysin makes chlorophyll detergent extractable

Detergent sensitivity of algal cells
(resistant to neutral, anionic detergents)

PBCV cell wall composition, New! - More compositions
- at least 8 sugars

PBCV whole virion lysin - intact virion lysins are host specific

Properties of ATCV WAL lysin
(lysin from lysed cells) - host nonspecific

New! - Properties of PBCV lysins

New! - pH and salt
dependence of PBCV1 WAL lysin

Calcium ion stabilizes WAL lysin

Effect of inhibitors on lysin
(sensitive to thiols, EDTA, proteases, resists iodoacetamide, peroxide, PMSF)

SAG WAL lysin acting on a substrate panel - substrate list
(substrates are fluorogenic)
Extracts have beta-NAc-glucosaminidase, alpha-glucosidase and beta-galactosidase activities
Activities of lysed cells - Glycosidases soluble, lysin in wall fraction

Chromatography of PBi soluble fractions, PBCV WAL lysin
(independent enzymes)

TCA precipitation of dansyl proteins
- Sarkosyl is an effective carrier

Host secretes NAc-beta-glucosaminidase
- part of autolysin?

Anion exchange chromatography of PBCV WAL lysin
- single component, weak negative charge

Cation exchange chromatography of PBCV WAL lysin - strong positive charge

Lysin assay of cloned A215L - Extracting cloned A215L

Extraction of lysin from PBCV with salt

Effect of alginate lyase on Chlorella strains

Lysin assay with Sybr Gold

Comparative cell specificities
of A215L, PBCV1 WAL lysin

Cell specificity of lysin dissociated from PBCV1 with LiCl

Chemical inhibtion of PBCV1 lysin (DEPC, DTT)

Cell specifity of ATCV lysin
is the same as that of PBCV1 lysin

Ammonium sulfate treatment of PBCV1

SDS gel of lysin prep

Preliminary secretome

Recent cation exchange

Cation exhange of lysin extracted from virions