Studies on a Lipoproteinase of Group a Streptococci

Krumwiede, Elma

doi:10.1084/jem.100.6.629

Cited by 33 publications

(30 citation statements)

References 7 publications

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“…The production of opacity in horse serum by group A streptococci is believed to be mediated by a lipoproteinase which acts upon the a,-lipoprotein fraction of the serum (Krumwiede, 1954;Rowen & Martin, 1963). Streptococci of groups other than A apparently do not produce opacity in serum, and within group A the serum opacity reaction (s.0.r.)…”

Section: Introductionmentioning

confidence: 99%

The Production of Opacity in Serum by Group A Streptococci and Its Relationship with the Presence of M Antigen

Widdowson

Maxted

Grant

1970

Journal of General Microbiology

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SUMMARYThe ability to produce opacity in horse serum is a characteristic of certain M types of group A streptococci. The types that produce the opacity factor (0 factor) are generally those for which it is difficult to produce good anti-M sera. M-positive (M+) and true M-negative (M-) variants of strains that belong to serotypes in which the serum opacity reaction (s.0.r.) is positive both possess the 0 factor, but there is a difference in binding of the factor to other cellular components in the two variants. The 0 factor is closely bound to the wall-membrane fraction of M-cells, whereas in M + cells it is easily extracted by Lancefield's method or I yo sodium deoxycholate.It is detectable in broth culture supernatants, in the cytoplasm and in the areas surrounding colonies in pour plates of M+ but not of M-cultures. The 0 factor is poorly antigenic, but when it is possible to obtain a good antiserum the inhibitory action is type specific. The 0 factor produced by several M types appears to be inhibited by normal rabbit serum. I N T R O D U C T I O NThe production of opacity in horse serum by group A streptococci is believed to be mediated by a lipoproteinase which acts upon the a,-lipoprotein fraction of the serum (Krumwiede, 1954;Rowen & Martin, 1963). Streptococci of groups other than A apparently do not produce opacity in serum, and within group A the serum opacity reaction (s.0.r.) is positive only in certain serotypes. Gooder (1961) and Kohler (1963) grew streptococci in a liquid medium consisting of 3 parts horse serum and I part Hartley digest broth, and recorded the presence of opacity in the culture supernatant as a positive s.0.r. Consistently s.0.r.-positive serotypes were those in which no M antigen was detectable, or for which satisfactory anti-M sera were difficult to prepare; on the other hand, members of a number of easily recognizable M serotypes were almost invariably s.0.r.-negative. Both workers suggested that there was an inverse relationship between a positive s.0.r. and the production of M antigen. Top & Wannamaker (1968a) incubated killed cells or deoxycholate extracts of fractions consisting predominantly of cell walls and cell membranes (' wall-membrane fractions') in horse serum. They confirmed that opacity was frequently produced by members of 'difficult' M types, but could find no true inverse relationship between M antigen and opacity production, except among type 1 2 strains.Most of the evidence concerning the supposed inverse relationship was obtained by examining collections of M-typable and M-untypable strains with common T-antigen

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Section: Introductionmentioning

confidence: 99%

The Production of Opacity in Serum by Group A Streptococci and Its Relationship with the Presence of M Antigen

Widdowson

Maxted

Grant

1970

Journal of General Microbiology

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“…Krumwiede (1954) and Rowen (1961) stated that in serum-free media the enzyme was cell bound with no extracellular activity. In initial studies with AN 124 we could find no extracellular enzyme activity, but subsequent studies have indicated that with certain strains some activity can be found in the supernatant, even when serum-free broth is used (L. W. Wannamaker, S. Skjold & F. H. Top, unpublished observations).…”

Section: Resultsmentioning

confidence: 99%

“…Extraction of the enzyme in a cell-free state Krumwiede (1954), Rowen (1961) and Rowen & Martin (1963) obtained the enzyme in a soluble form by extraction of whole cells with urea. In their hands this gives a product of high activity and considerable stability.…”

Section: Resultsmentioning

confidence: 99%

The serum opacity reaction ofStreptococcus pyogenes: general properties of the streptococcal factor and of the reaction in aged serum

Hill

Wannamaker

1968

J. Hyg.

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SUMMARYThe capacity of certain strains of Streptococcus pyogenes to produce opacity in aged horse serum has been studied. Cells from all stages of the growth cycle are able to produce opacity. Maximal activity is reached towards the end of the exponential phase of growth.Examination of cell fractions obtained by mechanical breakage and differential centrifugation suggested that the cell-bound activity is predominantly associated with the membrane fraction. Extraction with sodium deoxycholate yields a soluble fraction of high activity.There is considerable strain variation in heat stability of the serum opacity factor. Cell-bound activity is often quite resistant to heat, whereas extracted activity is less stable.Low concentrations of divalent cations have an activating effect, whereas high concentrations inhibit the serum opacity reaction. High concentrations of uni-valent cations are without effect on the cell-free enzyme but have an activating effect on the cell-bound enzyme.For both the cell-bound and the cell-free enzyme the pH optimum was 5·8.Although sensitive to trypsin and pepsin, the serum opacity factor appears to be resistant to streptococcal proteinase. Its activity is destroyed by formaldehyde and by periodate but is unaffected by a number of reducing agents.Pre-heating of the serum or the addition of iodoacetate did not affect the serum opacity reaction. The enhanced cholesterol esterification previously described with fresh serum appears to be a secondary reaction. Even when isolated by relatively gentle methods, α-lipoprotein serves as a substrate only in the presence of crystalline serum albumin.

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“…This serum opacity reaction (SOR) is mediated by a factor (OF) which acts upon the ccl-lipoprotein fraction of serum (Krumwiede, 1954).…”

Section: Introductionmentioning

confidence: 99%

Purification of the Extracellular Opacity Factor of a Strain of Group A Streptococcus M Type 2

Martinez¹,

Rippe

Fuller

et al. 1978

Journal of General Microbiology

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The extracellular Opacity Factor elaborated by a strain of group A streptococcus M type 2 was purified by ammonium sulphate precipitation, DEAE-cellulose and hydroxylapatite column chromatography and Sephadex G-200 gel filtration. Gel filtration experiments indicated that the Opacity Factor is constituted of high molecular weight proteins or protein aggregates which appear to dissociate into subunits of 66 000 minimum molecular weight as determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The purified preparations had no group A carbohydrate or T protein antigens. Passive haemagglutination and indirect immunofluorescence tests indicated that Opacity Factor is distinct from the M type 2 protein antigen.

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Studies on a Lipoproteinase of Group a Streptococci

Cited by 33 publications

References 7 publications

The Production of Opacity in Serum by Group A Streptococci and Its Relationship with the Presence of M Antigen

The Production of Opacity in Serum by Group A Streptococci and Its Relationship with the Presence of M Antigen

The serum opacity reaction ofStreptococcus pyogenes: general properties of the streptococcal factor and of the reaction in aged serum

Purification of the Extracellular Opacity Factor of a Strain of Group A Streptococcus M Type 2

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