PMID: 9523695Apr 2, 1998Paper

Isolation and characterisation of cAMP-dependent protein kinase from Candida albicans. Purification of the regulatory and catalytic subunits

European Journal of Biochemistry
Alicia ZeladaMaría Leonor Cantore

Abstract

cAMP-dependent protein kinase (PKA) from Candida albicans yeast cells was isolated and characterised. Structural parameters of the holoenzyme and those of its subunits suggested that C. albicans PKA is a tetramer of 287 kDa composed of two regulatory (R) subunits of 64 kDa and two catalytic (C) subunits of unusually large molecular mass of 78 kDa. The apparent Km for ATP and Kemptide were 30 microM and 60 microM respectively. The [A]0.5 for cAMP activation was 150 nM with a Hill coefficient of 1.6. The holoenzyme undergoes autophosphorylation on the R subunit, a characteristic of the type-II R subunits. Photoaffinity labeling with 8-azido-[32P]cAMP of crude extracts from yeast and mycelial cells strongly suggests that only one type of R subunit is present in the fungus. The R subunit was purified to apparent homogeneity as a protein of 64 kDa. A highly specific polyclonal antiserum raised against the purified protein immunoprecipitated a 64-kDa protein from crude extracts, indicating that the purified R subunit very probably represents the native form of the protein. The 78-kDa form of the C subunit was detected in crude extracts and in Mono Q Sepharose column fractions with heterologous anti-C Ig. It could be isolated by cAMP ...Continue Reading

Citations

Sep 25, 2012·Chemical Communications : Chem Comm·Gabrielle N Thibodeaux, Wilfred A van der Donk
Jan 29, 2000·Molecular Microbiology·A SonnebornJ F Ernst
Jan 13, 2006·Proceedings of the National Academy of Sciences of the United States of America·Andrew J PhillipsMark Ramsdale
Jan 17, 2009·Journal of Bioscience and Bioengineering·Yoshio KimuraKaoru Takegawa
Mar 3, 2009·Fungal Genetics and Biology : FG & B·José A Cervantes-ChávezJosé Ruiz-Herrera
Feb 6, 1999·The Journal of Biological Chemistry·S MilewskiG W Gooday
Mar 9, 2011·Archives of Biochemistry and Biophysics·Florencia KronbergSusana Passeron
Dec 27, 2011·Fungal Genetics and Biology : FG & B·Romina GiacomettiSusana Passeron

❮ Previous
Next ❯

Related Concepts

Related Feeds

Candidiasis

Candidiasis is a common fungal infection caused by Candida and it can affect many parts for the body including mucosal membranes as well as the gastrointestinal, urinary, and respiratory tracts. Here is the latest research.

Candidiasis (ASM)

Candidiasis is a common fungal infection caused by Candida and it can affect many parts for the body including mucosal membranes as well as the gastrointestinal, urinary, and respiratory tracts. Here is the latest research.

Candida albicans

Candida albicans is an opportunistic, fungal pathogen of humans that frequently causes superficial infections of oral and vaginal mucosal surfaces of debilitated and susceptible individuals. Discover the latest research on Candida albicans here.