Mammalian secreted group IIF phospholipase A2

Details Mammalian secreted group IIF phospholipase A2 Mammalian secreted group IIF phospholipase A2

2001-10-11

2004-11-02

Nashed, Nashaat T. (Department: 1652)

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Hydrolase

C435S198000, C424S094600, C536S023200

Reexamination Certificate

active

06812017

ABSTRACT:

FIELD OF THE INVENTION
This invention relates to DNA and peptide sequences encoding a novel mammalian secreted group IIF phospholipase A
2
and more particularly, a novel human group IIF phospholipase A
2
. The invention also relates to the use of this enzyme in methods for screening various chemical compounds.
BACKGROUND
Phospholipases A
2
(PLA
2
, EC 3.1.1.4.) form a superfamily of enzymes that catalyze the hydrolysis of glycerophospholipids at the sn-2 position, producing free fatty acids and lysophospholipids [1-4]. Many intracellular and secreted phospholipases A
2
(sPLA
2
s) have been cloned in recent years [2,5], and several of them are involved in a variety of physiological and pathological functions including lipid digestion, cell proliferation, production of lipid mediators of inflammation, antibacterial defense, and cancer [4,6].
Within the phospholipase A
2
superfamily, sPLA
2
s form a relatively homogenous family of enzymes. They are characterized by the presence of several disulfides, an overall conserved three-dimensional structure and a common Ca
2+
-dependent catalytic mechanism. Five novel mouse sPLA
2
s have been cloned during the last three years [7,8], and the mouse sPLA
2
family now comprises 8 distinct 14-16 kDa sPLA
2
s called group IB, IIA, IIC, IID, IIE, IIF, V and X, as well as otoconin-95, a sPLA
2
-like protein with peculiar structural properties [9,10]. Interestingly, genes for group IIA, IIC, IID, IIE, IIF, and V sPLA
2
s all map to mouse chromosome 4, suggesting the existence of a sPLA
2
gene cluster on this chromosome [8]. Group IB, IIA, IID, IIE, V and X sPLA
2
s, but not group IIF have been cloned from humans [11-13]. Conversely, group IIC sPLA
2
appears as a pseudogene in humans [14].
In addition, a novel human sPLA
2
with a predicted molecular mass of 55 kDA and a central domain similar to insect group III sPLA
2
s has recently been cloned [15], but it remains to be determined if this sPLA
2
is functional in the mouse species. This novel human sPLA
2
is also disclosed in the French International Patent Application N
o
01/59129. All mouse and human sPLA
2
s have distinct tissue distributions, suggesting that each of them exert non redundant functions that could be related to their different enzymatic properties [6,16,17], and/or their binding properties to specific receptors [17-19].
A comprehensive abbreviation system for the various sPLA
2
s is used thereafter: each sPLA
2
is abbreviated with a lowercase letter indicating the sPLA
2
species (m, h, for mouse and human, respectively) followed by capital characters identifying the sPLA
2
group (GI, GII, GIII, GV, and GX) and subgroup (A, B, C, D, E, F).
SUMMARY OF THE INVENTION
This invention relates to a mammalian secreted group IIF sPLA
2
which is Ca
2+
-dependent, maximally active at pH of about 7-8, and hydrolyzes phosphatidylglycerol versus phosphatidylcholine with about a 15-fold preference.


REFERENCES:
patent: WO 01/85956 (2001-11-01), None
Valentin et al. “Cloning and recombinant expression of human group IIF-secreted phospholipase A2” Biochem. Biophys. Res. Commu. 279, 223-228 (2000).*
Valentine et al. “On the diversity of secreted phospholipasees A2” J. Biol. Chem. 274, 31195-31202 (1999).

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