RESUMEN
Protein kinase CK2 is a ubiquitous serine/threonine kinase which is involved in many proliferation-related processes in the cell. It is composed of two regulatory beta-subunits and two catalytic alpha-subunits. Its regulation still remains mysterious in spite of many years of intense research. One of its regulators is the cdk inhibitory molecule p21(WAF1)-a protein which is expressed in situations of genotoxic stress. p21(WAF1) binds to the beta-subunit of CK2 and inhibits the activity of CK2. Using deletion mutants of CK2 beta as well as a peptide library consisting of 15-amino-acid-long peptides derived from the polypeptide chain of CK2 beta we mapped the binding region for p21(WAF1) on the polypeptide chain of CK2 beta. We localized an amino-terminal and a carboxy-terminal binding domain. Binding of p21(WAF1) to both regions of the CK2 beta-subunit interferes with the phosphotransferase activity of the CK2 holoenzyme.
Asunto(s)
Ciclinas/metabolismo , Proteínas Serina-Treonina Quinasas/química , Proteínas Serina-Treonina Quinasas/metabolismo , Secuencia de Aminoácidos , Animales , Sitios de Unión , Quinasa de la Caseína II , Inhibidor p21 de las Quinasas Dependientes de la Ciclina , Ciclinas/genética , Humanos , Técnicas In Vitro , Datos de Secuencia Molecular , Biblioteca de Péptidos , Mapeo Peptídico , Unión Proteica , Proteínas Serina-Treonina Quinasas/genética , Estructura Cuaternaria de Proteína , Estructura Terciaria de Proteína , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Eliminación de SecuenciaRESUMEN
Mdm2 is a cellular oncoprotein the most obvious function of which is the down-regulation of the growth suppressor protein p53. It represents a highly phosphorylated protein but only little is yet known about the sites phosphorylated in vivo, the kinases that are responsible for the phosphorylation or the functional relevance of the phosphorylation status. Recently, we have shown that mdm2 is a good substrate for protein kinase CK2 at least in vitro. Computer analysis of the primary amino acid sequence of mdm2 revealed 19 putative CK2 phosphorylation sites. By using deletion mutants of mdm2 and a peptide library we identified the serine residue at position 269 which lies within a canonical CK2 consensus sequence (EGQELSDEDDE) as the most important CK2 phosphorylation site. Moreover, by using the mdm2 S269A mutant for in vitro phosphorylation assays this site was shown to be phosphorylated by CK2. Binding studies revealed that phosphorylation of mdm2 at S269 does not have any influence on the binding of p53 to mdm2.
Asunto(s)
Proteínas Nucleares , Proteínas Serina-Treonina Quinasas/química , Proteínas Proto-Oncogénicas/química , Secuencia de Aminoácidos , Animales , Sitios de Unión , Western Blotting , Quinasa de la Caseína II , Línea Celular , Codón , Electroforesis en Gel de Poliacrilamida , Escherichia coli/metabolismo , Humanos , Insectos , Datos de Secuencia Molecular , Mutación , Péptidos/química , Fosforilación , Plásmidos/metabolismo , Unión Proteica , Estructura Terciaria de Proteína , Proteínas Proto-Oncogénicas/metabolismo , Proteínas Proto-Oncogénicas c-mdm2 , Proteínas Recombinantes/metabolismo , Homología de Secuencia de Aminoácido , Proteína p53 Supresora de Tumor/metabolismoRESUMEN
p53 is one of the most powerful negative regulators of growth. To manage this in an efficient way it has to interact with a set of different cellular proteins. Most contacts with the cellular environment occur in the N- or the C-terminal domain of the protein. Since we previously found that p53 binds to the regulatory beta-subunit of CK2 we now analyzed N- and C-terminal domains of p53 separately for the binding of protein kinase CK2, an enzyme which seems to have a certain importance for proliferation processes. With different overlay assays we could map the binding domain of protein kinase CK2 to a sequence between amino acids 325-344, a region which coincides with the interaction domain of some other p53 binding proteins. We also found that the regulatory beta-subunit of protein kinase CK2 binds independent of the catalytic alpha-subunit to this C-terminal domain of p53.
Asunto(s)
Proteínas Serina-Treonina Quinasas/metabolismo , Proteína p53 Supresora de Tumor/metabolismo , Secuencia de Aminoácidos , Unión Competitiva , Quinasa de la Caseína II , Dominio Catalítico , Humanos , Datos de Secuencia Molecular , Mapeo Peptídico , Unión Proteica , Proteína p53 Supresora de Tumor/químicaRESUMEN
Medically fragile infants and children present a host of challenges. The neonatal intensive care unit (NICU) is initially the mainstay of care for these infants, but it does not meet the critical developmental interactive needs of the child and parents to ensure positive psychosocial bonding. The care of these infants is further complicated by high daily costs (ranging from $1,000 to $2,500 per neonatal intensive care bed day) and lengthy, expensive inpatient stays (typically ranging from one week to five months or more). FHP Utah, a managed care organization, met these challenges through an innovative case management-oriented home care program called "Welcome Home." The program has produced a major impact on the quality and the cost of care for medically fragile infants. This article summarizes the first year's experience in implementing and operating the program.
Asunto(s)
Servicios de Atención de Salud a Domicilio/organización & administración , Cuidado Intensivo Neonatal/organización & administración , Relaciones Padres-Hijo , Atención Progresiva al Paciente/organización & administración , Humanos , Recién Nacido de Bajo Peso , Recién Nacido , Desarrollo de Programa , UtahRESUMEN
The evidence is overwhelming that sharp reductions in health care costs can be achieved by reducing or eliminating reliance on expensive hospital care. In surgery, this is not always possible. However, significant savings can be achieved through reducing the length of time that a surgery patient must spend in the hospital. In outpatient surgery, the hospital stay is reduced to a single day. In this article, the authors describe a home surgical recovery program that has had great success in reducing hospital stays for even more complicated surgical procedures, frequently matching the achievements of outpatient surgery programs.
Asunto(s)
Servicios de Atención de Salud a Domicilio/normas , Cuidados Posoperatorios/normas , Adulto , Anciano , Procedimientos Quirúrgicos Ambulatorios/normas , California , Humanos , Persona de Mediana Edad , Satisfacción del Paciente/estadística & datos numéricos , Evaluación de Programas y Proyectos de Salud/estadística & datos numéricos , Encuestas y CuestionariosRESUMEN
Peter R. Scholtes has a unique perspective on what it takes to build a world-class quality organization: A transformation of the relationships, environment, and dynamics within and between individuals and groups throughout an organization. He brings an organizational development perspective to quality management and, in particular, to the approach and practices advocated by W. Edwards Deming. This interview explores Mr. Scholtes' in-depth understanding and sometimes controversial views on quality improvement teams, team training, and performance appraisal.