title: Catalytic signature of a heat-stable, chimeric human alkaline phosphatase with therapeutic potential.
abstract: Recombinant alkaline phosphatases are becoming promising protein therapeutics to prevent skeletal mineralization defects, inflammatory bowel diseases, and treat acute kidney injury. By substituting the flexible crown domain of human intestinal alkaline phosphatase (IAP) with that of the human placental isozyme (PLAP) we generated a chimeric enzyme (ChimAP) that retains the structural folding of IAP, but displays greatly increased stability, active site Zn²⁺ binding, increased transphosphorylation, a higher turnover number and narrower substrate specificity, with comparable selectivity for bacterial lipopolysaccharide (LPS), than the parent IAP isozyme. ChimAP shows promise as a protein therapeutic for indications such as inflammatory bowel diseases, gut dysbioses and acute kidney injury.
authors.affiliation: Sanford Children's Health Research Center, Sanford-Burnham Medical Research Institute, La Jolla, California, United States of America.
authors.forename: Tina
authors.initials: T
authors.lastname: Kiffer-Moreira
authors.affiliation: Sanford Children's Health Research Center, Sanford-Burnham Medical Research Institute, La Jolla, California, United States of America.
authors.forename: Campbell R
authors.initials: CR
authors.lastname: Sheen
authors.affiliation: Sanford Children's Health Research Center, Sanford-Burnham Medical Research Institute, La Jolla, California, United States of America.
authors.forename: Kellen Cristina da Silva
authors.initials: KC
authors.lastname: Gasque
authors.affiliation: Departamento de Química, FFCLRP-USP, Ribeirão Preto, São Paulo, Brazil.
authors.forename: Mayte
authors.initials: M
authors.lastname: Bolean
authors.affiliation: Departamento de Química, FFCLRP-USP, Ribeirão Preto, São Paulo, Brazil.
authors.forename: Pietro
authors.initials: P
authors.lastname: Ciancaglini
authors.affiliation: AM-Pharma BV, Bunnik, The Netherlands.
authors.forename: Andrea
authors.initials: A
authors.lastname: van Elsas
authors.affiliation: Department of Cardiovascular Sciences, Center for Molecular and Vascular Biology, University of Leuven, Leuven, Belgium.
authors.forename: Marc F
authors.initials: MF
authors.lastname: Hoylaerts
authors.affiliation: Sanford Children's Health Research Center, Sanford-Burnham Medical Research Institute, La Jolla, California, United States of America.
authors.forename: José Luis
authors.initials: JL
authors.lastname: Millán
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism*; D000595:Amino Acid Sequence; D002384:Catalysis; D020134:Catalytic Domain; D005260:Female; D058851:GPI-Linked Proteins / Q000378:metabolism; D006358:Hot Temperature; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D008969:Molecular Sequence Data; D010920:Placenta / Q000378:metabolism; D011247:Pregnancy; D017510:Protein Folding; D055550:Protein Stability; D011994:Recombinant Proteins / Q000378:metabolism*
chemical_list: D058851:GPI-Linked Proteins; D007527:Isoenzymes; D011994:Recombinant Proteins; C063088:ALPI protein, human; D000469:Alkaline Phosphatase