title: A missense mutation Pro157 Arg in lipoprotein lipase (LPLNijmegen) resulting in loss of catalytic activity.
abstract: Here we report on the molecular defect that leads to a deficiency of lipoprotein lipase (LPL) activity in a proband of Dutch descent. Southern-blot analysis of the LPL gene from the patient did not reveal any major DNA rearrangements. Sequencing of polymerase-chain-reaction-amplified DNA revealed that the proband is a homozygote for G725C, resulting in a substitution of Pro157 for Arg. This substitution alters a restriction site for PvuII, which allowed rapid identification of the mutant allele in family members. Site-directed mutagenesis and transient expression of the mutant LPL in COS cells produced an enzymatically inactive protein, establishing the functional significance of this mutation. This naturally occurring mutation which alters the Pro157 adjacent to Asp156 of the proposed catalytic triad, indicates that this region of the protein is indeed crucial for LPL catalytic activity.
authors.affiliation: Centre for Hemostasis, Thrombosis, Atherosclerosis and Inflammation Research, University of Amsterdam, The Netherlands.
authors.forename: T
authors.initials: T
authors.lastname: Bruin
authors.forename: J J
authors.initials: JJ
authors.lastname: Kastelein
authors.forename: D E
authors.initials: DE
authors.lastname: Van Diermen
authors.forename: Y
authors.initials: Y
authors.lastname: Ma
authors.forename: H E
authors.initials: HE
authors.lastname: Henderson
authors.forename: P M
authors.initials: PM
authors.lastname: Stuyt
authors.forename: A F
authors.initials: AF
authors.lastname: Stalenhoef
authors.forename: A
authors.initials: A
authors.lastname: Sturk
authors.forename: J D
authors.initials: JD
authors.lastname: Brunzell
authors.forename: M R
authors.initials: MR
authors.lastname: Hayden
mesh_terms: D000328:Adult; D000595:Amino Acid Sequence; D001120:Arginine / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D002384:Catalysis; D002460:Cell Line; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D005260:Female; D006801:Humans; D008072:Hyperlipoproteinemia Type I / Q000235:genetics*; D008071:Lipoprotein Lipase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010375:Pedigree; D016133:Polymerase Chain Reaction; D011392:Proline / Q000235:genetics*; D013329:Structure-Activity Relationship; D014162:Transfection
chemical_list: D001120:Arginine; D011392:Proline; D008071:Lipoprotein Lipase; C025630:CAGCTG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific